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Recaldent

TM

(CPP-ACP)

Literature Overview
G C M I P a s t e TM – A b s t r a c t B o o k l e t

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GC MI Paste™ / GC MI Paste Plus™ and GC Tooth Mousse® and GC Tooth Mousse Plus® are synonymous. In the United States it is called MI Paste or MI Paste Plus. In Europe and Australia it is called Tooth Mousse or Tooth Mousse Plus. 10/2007

GC MI Paste – Abstract booklet – October 2007

Literature overview Recaldent
1. 2. 3. 4. 5. 6. 7. 8. 9.

TM

(CPP-ACP)

Dental caries in the cotton rat. VI. The effect of the amount of protein, fat and carbohydrate in the diet on the incidence and extent of carious lesions. Schweigert BS, Shaw JH, Zepplin M, Elvehjem CA. J Nutr 31: 439-447 (1946) Effects of dietary composition on tooth decay in the albino rat. Shaw JH. J Nutr 41: 23-23 (1950) Protein factors and experimental rat caries. Bavetta LA, McClure FJ. Nutr. 63: 107 -117 (1957) Calcium phosphate sequestering phosphopeptide from casein. Reeves RE, Latour NG. Science 128: 472 (1958) Effects of various sucrose-casein ratios in purified diets on the teeth and supporting structures of rats. Holloway PJ, Shaw JH, Sweeny EA. Arch Oral Biol 3: 185-200 (1961) A review of the effect of milk on dental caries. Reynolds EC, Storey E. Aust J Dairy Tech 34:175-179 (1979) Effect of milk on caries incidence and bacterial composition of dental plaque in the rat. Reynolds EC, Johnson IH. Arch Oral Biol 1981 26:5 445-51 Phosphoprotein inhibition of hydroxyapatite dissolution. Reynolds EC, Riley PF, Storey E. Calcif Tissue Int 1982 34 Suppl 2: S52-6 Effect of adsorbed protein on hydroxyapatite zeta potential and Streptococcus mutans adherence. Reynolds EC, Wong A. Infect Immun 1983 Mar 39:31 285-90

10. Effect of casein and whey-protein solutions on caries experience and feeding patterns of the rat. Reynolds EC, del Rio A. Arch Oral Biol 1984 29:11 927-33 11. Effect of cheese, with and without sucrose, on dental caries and recovery of streptococcus mutans in rats. Rosen S, Min DB, Herper DS, Herper WJ, Beck EX, Beck FM. J Dent Res 63: 894-896 (1984) 12. Cariostatic evaluation of cheeses with diverse physical and compositional characteristics. Harper DS, Osborn JC, Clayton R, Hefferren JJ. Caries Res 20:123-130 (1986) 13. Role of streptococcus mutans in human dental decay Loesche WJ. Microbiol Rev 1986 Dec;50(4): 353-380 14. Modification of food cariogenicity in rats by mineral-rich concentrates from milk. Harper DS, Osborn JC, Clayton R, Hefferren JJ. J Dent Res 66:42-45 (1987) 15. Effects of water soluble components of cheese on experimental caries in humans. Silva MfdeA, Burgess RC, Sandham HJ, Jenkins GN. J Dent Res 66: 38-41 (1987) 16. The prevention of sub-surface demineralization of bovine enamel and change in plaque composition by casein in an intra-oral model. Reynolds EC. J Dent Res 1987 Jun 66:6 1120-7 17. Reduction of chocolate's cariogenicity by supplementation with sodium caseinate. Reynolds EC, Black CL. Caries Res 1987 21:5 445-51 18. Confectionery composition and rat caries. Reynolds EC, Black CL. Caries Res 1987 21:6 538-45 19. The effects of cheese snacks on caries in desalivated rats. Krobicka A, Bowen WH, Pearson S, Youn DA. J Dent Res 66: 1116-1119 (1987) 20. Protein dissimilation by human salivary-sediment bacteria. Reynolds EC, Riley PF. J Dent Res 68: 124-129 (1989) 21. Cariogenicity of a confection supplemented with sodium caseinate at a palatable level. Reynolds EC, Black CL. Caries Res 23:368370 (1989) 22. A 24-month clinical study of the incidence and progression of dental caries in relation to consumption of chewing gum containing xylitol in school preventive programs. Kandelman D, Gagnon G. J Dent Res 69(11): 1771-1775 (1990) 23. Efficient solution-phase synthesis of multiple O-phosphoseryl-containing peptides related to casein and statherin. Perich JW, Kelly DP, Reynolds EC. Int J Pept Protein Res 1992 Aug 40:2 81-8 24. The use of synthetic phosphopeptides for epitope mapping of the αs1- casein phosphopeptide segment 59 -79. Perich JW, Reynolds EC. Bioorg Med.Chem Lett 1992 2:1153 – 1154 25. An in situ model for simultaneous assessment of inhibition of demineralization and enhancement of remineralization. Featherstone JDB, Zero DT. J Dent Res 71(Spec. Iss.): 804-810 (1992) 26. The analysis of multiple phosphoseryl-containing casein peptides using capillary zone electrophoresis. Reynolds EC. J Chromatogr 1993 Sep 3 646:2 391-6 Adamson N, Riley PF,

27. A selective precipitation purification procedure for multiple phosphoseryl-containing peptides and methods for their identification. Reynolds EC, Riley PF, Adamson NJ. Anal Biochem 1994 Mar 217:2 277-84

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GC MI Paste – Abstract booklet – October 2007

28. High performance capillary electrophoresis of casein phosphopeptides containing 2-5 phosphoseryl residues: relationship between absolute electrophoretic mobility and peptide charge and size. Adamson NJ, Reynolds EC. Electrophoresis 16: 525-528 (1995) 29. A 1H-NMR study of the casein phosphopeptide alpha s1-casein(59-79). Huq NL, Cross KJ, Reynolds EC. Biochim Biophys Acta 1995 Mar 15 1247:2 201-8 30. Anticariogenicity of calcium phosphate complexes of tryptic casein phosphopeptides in the rat. Reynolds EC, Cain CJ, Webber FL, Black CL, Riley PF, Johnson IH, Perich JW. J Dent Res 1995 Jun 74:6 1272-9 31. In situ caries models. Zero DT. Adv Dent Res 9(3): 214-230 (1995) 32. Characterisation of tryptic casein phosphopeptides prepared under industrially-relevant conditions. Adamson NJ, Reynolds EC. Biotec Bioeng 1995 45:196-204 33. Role of models in assessing new agents for caries prevention--non-fluoride systems. Roberts AJ. Adv Dent Res 1995 Nov;9(3):30411; discussion 312-4 34. Incorporation of caseinoglycomacropeptide and caseinophosphopeptide into the salivary pellicle inhibits adherence of mutans streptococci. Schupbach P, Neeser JR, Golliard M, Rouvet M and Guggenheim B. J Dent Res, Vol 75, 1779-1788 (1996) 35. Ability of a b-casein phosphopeptide to modulate the precipitation of calcium phosphate by forming amorphous dicalcium phosphate nanoclusters Holt C, Wahlgren NM and Drakenberg T. Biochem. J (1996) 314 (1035–1039) 36. Dairy products and dental health. Reynolds EC. Proc Nutr Soc Aus 1997 76:1587-1595 37. Remineralization of enamel subsurface lesions by casein phosphopeptide-stabilized calcium phosphate solutions. Reynolds EC. J Dent Res 1997 Sep 76:9 1587-95 38. Anticariogenic complexes of amorphous calcium phosphate stabilized by casein phosphopeptides: a review. Reynolds EC. Spec Care Dentist 1998 Jan-Feb 18:1 8-16 39. Anticariogenic casein phosphopeptides. Reynolds EC. Prot Peptide Lett 1999 295-303 40. Advances in enamel remineralization: anticariogenic casein phosphopeptide-amorphous calcium phosphate. Reynolds EC, Black CL, Cross KJ, Eakins D, Huq NL, Morgan MV, Nowicki A, Perich JW, Riley PF, Shen P, Talbo G, Webber FW J Clin Dent 1999 X(2):86-88 41. Calcium phosphates in demineralization/remineralization processes. Legeros RZ J Clin Dent 1999 X:65-73 42. The role of phosphopeptides in caries prevention. Reynolds EC. Dental Perspectives 1999 3:6-7 43. Molecular modelling of a multiphosphorylated sequence motif bound to hydroxyapatite surfaces. Huq NL, Cross KJ, Reynolds EC. J Mol Mode 2000 6:35-47 44. The use of casein phosphopeptides in oral care products for the prevention and treatment of early enamel caries. Reynolds EC. Aust J Dairy Technol 2000 55:1-6 (not published) 45. Binding characteristics of streptococcus mutans for calcium and casein phosphopeptide. Rose RK. Caries Res 2000 34 427-431 46. Remineralisation of fluorotic enamel lesions by casein phosphopeptide – amorphous calcium fluorophosphate (CPP-ACFP) solution. Roberts MJ, Messer LB, Reynolds EC. IADR,ANZ division, Abstract 54,2000 47. Experimental study of phosphopeptide in promoting tooth remineralization Zhang L, Li Z, Dong Y. Chin J Dent Res 2000 May;3(1):27-30 48. Effects of an anticariogenic casein phosphopeptide on calcium diffusion in streptococcal model dental plaques. Rose RK. Arch Oral Biol, Volume 45, Issue 7, 2000, Pages 569-575 49. Enamel remineralization by chewing gum containing casein phosphopeptide - amorphous calcium phosphate. Reynolds EC, Shen P, Cai F, Nowicki A, Vincent J. IADR,General session,Chiba,Abstract 0489,2001 50. Structural studies of the ß - casein phosphopeptide bound to amorphous calcium phosphate. Cross KJ, Huq NL, Eakins D, Reynolds EC. IADR,General session,Chiba,Abstract 0490,2001 51. Ultrastructural studies of the casein phosphopeptide- amorphous calcium phosphate nanoclusters. Cross KJ, Huq NL, Eakins D, Reynolds EC. IADR,General session,Chiba,Abstract 0491,2001 52. Cation-dependent structural features of beta-casein-(1-25). Cross KJ, Huq NL, Bicknell W, Reynolds EC. Biochem J 2001 May 15 356:Pt 1 277-86 53. MALDI-PSD-MS analysis of the phosphorylation sites of caseinomacropeptide. Talbo GH, Suckau D, Malkoski M, Reynolds EC. Peptides 2001 Jul 22:7 1093-8 54. Health aspects of dairy products- Dairy products in relation to caries prevention and oral health. Invited review. Encyclopedia of Dairy Sciences 2001 Reynolds EC.

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Holler BE. Yoshii E. Walker G. Honolulu 2004 . Koganei M. Accepted for publication. Remineralization of enamel subsurface lesions in situ by sugar-free lozenges containing casein phosphopeptide-amorphous calcium phosphate. De Munck J. Australian Dental Journal ADRF Special Research Supplement 2004. Cai F. EADR Istanbul. Morita M. Hinoura K. Aust J Dairy Tech 58:79-81 (2003) 65. Sakaguchi Y. Honolulu 2004 . Morgan MV. Fluoride effect on acid resistance capacity of CPP-ACP containing material. Paediatric Dent. Honolulu 2004 . Ikeda T. Takamizawa T. Inage H.General session. 78. Acid Resistance of Enamel Subsurface Lesions Remineralized by a Sugar-Free Chewing Gum Containing Casein PhosphopeptideAmorphous Calcium Phosphate. Cai F. Yamanaka K. Friedl K-H. Reynolds EC. Honolulu 2004 . Sum M.261-275 79. Erosion of human dental enamel by sports drinks. IADR. Reynolds EC.Abstract 2049 71. nd IADR. Reynolds EC. USA / Caries Res 2005. Shen P. Effects of cheese and milk containing CPP-ACP on enamel remineralization. Reynolds EC.93:271-5 59. Cai F. Jung H. Mazzaoui SA. 2004 pp. 82 General Session.amorphous calcium phosphate (CPP – ACP).General session. Shen P. Retention in plaque and remineralization of enamel lesions by various forms of calcium in a mouthrinse or sugar-free chewing gum. J Dent Res 2001 Dec 80:12 2066-70 56. Shen P. January 2004 68. Remineralization power by xylitol chewing gums. Matsuda Y. Dent Practice 2001 November / December 57.Abstract 2997 72. Poitevin A. Caries prevention potential of a tooth-coating material containing casein phosphopeptide . IADR. Miyazaki M. Aus Dent J 2003 48:4 240-243 63. phosphate and fluoride delivery vehicle – the multiphosphorylated peptide alpha s1-casein (59-79) complexed with amorphous calcium fluoride phosphate. Inaba D. Reynolds EC. Coutinho E. Miyake R. Stanton D. Minami K. Caries Res 2004. Nowicki A. Ramalingam L. Walker G. Cai F. nd Cross KJ. 82 General Session. IADR. Hiller KA. Tanaka T. Textbook: Preservation and Restoration of Teeth 2nd edition 2005. Abstract 94 – 52 ORCA Congress. Dashper SG. Yonemitsu M. Reynolds EC. Walsh LJ. NMR studies of a novel calcium. Reynolds EC. Van Meerbeek B. Reynolds EC. nd IADR. Using ultrasound transmission velocity to analyze demineralization of tooth substrate. Cai F. Huq NL. Cross KJ. Cai F.Abstract 2045 nd 69. J Dent Res 2003 Nov 82:11 914-8 60. Enamel remineralization by a mouthrinse containing casein phosphopeptide-amorphous calcium phosphate and fluoride in an in situ model. Molecular of anticariogenic casein phosphopeptide aS2-CN(2-20) NMR spectroscopy derived constraints. Additional aids to the remineralisation of tooth structure. IADR. 82 General Session.38:551556 67.2003 61. Yamaguchi K. Japan J Conserv Dent 2004 Jun 47 Spring Issue 24 – Abstract B-4 73. Sano H. July 2005. Takamizawa T. 82 General Session. La reminéralisation des lesions carieuses (2) synergies thérapeutiques / The remineralisation of caries lesions: joint therapies.Abstract 1007. Yoshii E. Acid resistance of enamel subsurface lesions remineralized by a sugar-free chewing gum containing casein phosphopeptideamorphous calcium phosphate (CPP-ACP). Reynolds EC. Braem M. Komatsu H. Reynolds EC. Deangelis EF.GC MI Paste – Abstract booklet – October 2007 55. A clinical trial of the anticaries efficacy of casein derivatives complexed with calcium phosphate in patients with salivary gland dysfunction. Reynolds C. Australian Dental Journal ADRF Special Research Supplement 2004. Reynolds EC. Walker G.15 n°3. Remineralization of early enamel caries by anticariogenic casein phosphopeptide-amorphous calcium phosphate nanocomplexes. Tyas MJ. Hinoura K. 39:319 80. Ramalingam L . An in vitro investigation of the effects of casein phosphopeptide-stabilized amorphous calcium phosphate (CPP-ACP) on erosion of human dental enamel by a sports drink. Reynolds EC. Reynolds C. Realites Cliniques Vol. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2002. Reynolds C. Vincent J. Cross KJ. Caries Res 2004. Walker GD. Determination of demineralization of tooth substrate by use of an ultrasonic device. Molecular modelling of the multiphosphorylated casein phosphopeptide alpha s1-casein (59-79). J Dairy Res 71:1-5 (2004) 66. Dairy components in oral health.San Diego 2002 . Iijima Y. Sato T. 25-28 August 2004 – Abstract 0136 74. Yamaguchi K. Van Landuyt K. Reynolds EC. Accepted for publication 64. Fluoride uptake and distribution in enamel and dentin after application of different fluoride solutions. Kariya S. Oda M. Eakins D. Miller C. Huq NL. Reynolds EC Biomaterials. Ung M. Shen P. Lasfargues JJ. Martin JM. Huq NL. Shen P. Miyazaki M. Messer LB. Investigation of the binding of casein phosphopeptides to the major enamel pellicle proteins. Messer LB. Incorporation of Casein Phosphopeptide-Amorphous Calcium Phosphate into a Glass-ionomer Cement.Abstract 2046 70. Ando S. 49:4 75. Murata Y. Cross KJ. Clinical effectiveness of a CPP-ACP crème for tooth hypersensitivity treatment. Reynolds EC. Hay KD and Thomson WM. Reynolds EC. Burrow MF. J Dent Res 2003 Mar 82:3 206-11 62. Indianapolis. Schmalz G. Huq NL. Remineralization of enamel subsurface lesions by sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate. Shen P. p111-118 77. Sato T. Clin Oral Invest (2002) 6:137-144 4 . Peumans M.Goteborg. Iijima Y. 49:4 76.Abstract 2810 58.38:551-556. nd Inage H.

Takamizawa. Sakaguchi Y. * 98. Kurokawa. P. Cai F. F. F. Brisbane. Australia. 28 June .G. Australia. Abstract 0654 . Morgan.72. Manton. K. Australia. Kato S. Ranjitkar.C. * 96. Brisbane.C. th Reynolds EC. Bailey. G. Australia. * 92. JCDA. Kato S. M. Al-Zraikat. Reynolds. G-H. B.C.1 July 2006. M. Walker GD. Messer.1 July 2006.5 * 85. Yuan Y.H.1 July 2006. Miyazaki. June 2006. S. Kaidonis. M.V. Palamara. Moule. 83 General Session. Poolthong. Cai.L. F. E.S. Brisbane. Panich.C. Cochrane NJ. Australia. 28 June . Plaque microcosm biofilm mineralization by CPP-ACP and calcium-phosphate-monofluorophosphate-urea mineralizing solution. Reynolds C. Sukasaem. J. 28 June .1 July 2006. Australia. E. Abstract 185 – 84 General Session of the IADR. T. L. Abstract th 2424 – 84 General Session of the IADR. Kariya S.D. N. Ranjitkar.. Messer LB. Sudjalim. Tran KK.1 July 2006. Shen. Caries Res 2007. Manton. 4 :15-21 * 84. Morgan. Australia. P.1 July 2006. G.1 July 2006. 28 June . Brisbane. A chemical approach. Cakar A. th Walker.F. Wang W. Reynolds. Vol. CPP-ACP gum slows progression and enhances regression of dental caries. Australian Dental Journal 2006. D. Wilson. 28 June . Chen L.84 General Session of the IADR. No. Australia. J. D. Cai. Shen P. Casein phosphopeptide-amorphous calcium phosphate paste: root surface caries formation. E. Shen P. Cross. 28 June . M. T.G.C. Acid resistance of remineralized enamel by a sugar-free chewing gum. Abstract 190 – 84 General Session of the IADR.Abstract 2055 82. Sato T. G. Wong. QLF and TMR analysis of CPP-ACFP remineralized enamel in vitro. H. L. Effect of CPP-ACP on hardness of enamel eroded by Cola-drink. Inage. Tsao. * 90.1 July 2006. Richards. T. N. Palamara. Reynolds EC. 28 June . C. Brisbane. Walker GD. J. Effect of CPP-ACP versus potassium nitrate on cervical dentinal hypersensitivity. Shen P. 83 General Session. * 89. th Adams. * 93. D. 2:133-137 Angelis. Australia. Minimal intervention dentistry: part 1. L. 28 June . 230-236* 105. Remineralization by chewing gum containing CPP-ACP and citric acid.L. Australia.K. L. M. Strategies for addressing the new caries challenge in older patients. Abstract 0020 – 45 Annual Meeting of Australian/New Zealand Division of the IADR. M. Ingenito A. IADR. Yamaguchi. Baltimore 2005 . S. Remineralization potential of CPP-ACP and its synergy with fluoride.84 General Session of the IADR. 52 5 . Prevenzione Odontostomatologica 2005 . th Burrow. Foo. Woods. Brisbane.C.51 :(4):284-289 * 101.R. Sissons. Burrow. Reynolds C. Chalmers JM. Flaitz C. Baltimore 2005 . Enamel wear prevention under conditions simulating bruxism and acid regurgitation. nd IADR.1 July 2006. D. Abstract 1673 – 84 General Session of the IADR. M. Sato T. Structure and 15N-Dynamics of casein phosphopeptide-amorphous calcium phosphate nanocomplexes. * 94. Reynolds EC. Reynolds. Abstract 2445 – 84 General Session of the IADR.5. Cai F. Prevention of white spot lesions in orthodontic practice: a contemporary review. * th th th nd 95. 103. Australia. K. Manton DJ. Abstract 947 – 84 General Session of the IADR. Brisbane. Kariya S. * 99. C. th Reynolds EC. Townsend. T. Moore.1 July 2006. Brisbane.R. 2006 86. Brisbane. R. * 97. 28 June . Effect of CPP-ACP paste on mechanical properties of bovine enamel as determined by an ultrasonic device. G. 25-28 September 2005 102. M. Brisbane. P. Townsend. Reynolds. S. Brisbane. Brisbane. H. Abstract 2534 – 84 General Session of the IADR. Australia. S. An in vitro study of wear prevention in dentine.1 July 2006. Manton DJ. Abstract 192 – 84 General Session of the IADR. * 88. th Abstract 191 – 84 General Session of the IADR. S.S. Le. Australian Dental Journal 2007. Cai F. Abstract th 189 – 84 General Session of the IADR.J. 28 June . 104. Resin bonding using an all-etch or self-etch adhesive to enamel after carbamide peroxide and/or CPP-ACP treatment. C. Walker.A. th Richards. In situ remineralisation by sugar-free gums.B. M. Brisbane. * 91. Chen L. Abstract 1269 – 84 General Session of the IADR. K. J. E. Narayana. C. one containing CPP-ACP. Australia.C. 41:204-207 106. Journal of Dentistry (2006) 34. Adams. Nuove strategie nella prevenzione della carie dentaria:studio sperimentale sui caseino-fosfopeptidi.J. Sangianantoni G. G. C. th Reynolds.C. Reynolds.1 July 2006. Iijima Y. Thomas. Abstract 2424 – 84 General Session of the IADR.D. Remineralization of white spot lesions in situ by tooth mousse.J. Malipatil.1 July 2006.J. Brisbane. Shen. th Wong. th Huq. Kaidonis. Brisbane.1 July 2006.A. Hicks J. 28 June .1 July 2006. Abstract 184 – 84 General Session of the IADR.E. Preventing acid induced enamel demineralization using CPP-ACP containing paste. Incorporation of casein phosphopeptide-amorphous calcium phosphate into a temporary cement. Australia. EAPD. Brisbane. Cai F. Messer. Reynolds C. Sakaguchi Y. Australia. th Abstract 0653 . Cochrane NJ. Walsh LJ. Takamizawa.A. Miyazaki. 87. 28 June . H. Abstract 2538 – 84 General Session of the IADR. H. Yamaguchi. N. T. Remineralization by a mouthrinse containing CPP-ACP at pH 5. th Hall A. Troianiello S. Devani C. Incorporation of casein-phosphopeptide-amorphous calcium phosphate into glass ionomer cement. 28 June . * 100. Ferrazzano GF.Abstract 3275 83. Attard. Kim. Iijima S. H.GC MI Paste – Abstract booklet – October 2007 81.J. Nagao S. Australia. Improved plaque uptake and enamel remineralization by fluoride with CPP-ACP. 28 June . G. Cochrane.1 July 2006.V. Ultrasonic determination of the effect of casein phosphopeptide-amorphous calcium phosphate paste on the demineralization of bovine dentin. Nishimura M. Inage. Manton DJ. 28 June . Promoting remineralization: using casein phosphopeptide – stabilized amorphous calcium (fluoride) phosphate. H. Cochrane. 28 June . . Amsterdam 8-11 June.

May 2007. Effect of a CPP-ACP agent on the demineralization and remineralization of dentine in vitro. Inage.J. M. Milnar. Vougiouklakis. Vol. No. Sakaguchi. Irokawa. C.28(5):234-240 6 . T. T. G. 115-120. L. Remineralization of enamel lesion by a novel cream with both CPP-ACP and fluoride. F. H. Effect of CPP-ACP paste on tooth mineralization: an FE-SEM study. M.GC MI Paste – Abstract booklet – October 2007 107. S. K. S. Miyazaki. Kariya. S. Journal of oral Science. Iijima. A. Y. Compendium. Chen Y. Takamizawa. T.49. Yamaguchi. Poster session 136 – 54th Annual ORCA Congress 2007 110. Sato. 2007 108. Watanabe. Considering biomodification and remineralization techniques as adjuncts to vital tooth-bleaching regimens. Rahiotis. Ando. Kato. Journal of Dentistry 35 (2007) 695-698 109. Oshiro.2.

The incidence and extent of tooth decay were low when a ration approximating milk solids in composition was fed. Shaw JH. In each of the above results. 2. Dental caries in the cotton rat. Caloric restriction resulted in appreciably less tooth decay than in the control rats which were allowed to eat ad libitum.GC MI Paste – Abstract booklet – October 2007 1. the influence of dietary variation on the initiation and development of carious lesions in the caries-susceptible white rat is extremely comparable to the effect obtained with the same regimens in cotton rats. 7 . unlike its strong influence in the cotton rat. When 50 parts of casein and 10 parts of lard were fed. Administration of either mineralized fresh or evaporated milk as the sole source of nutrients resulted in extremely low degrees of dental decay. The isocaloric substitution of fat and protein for part of the sucrose in the purified ration resulted in substantial reductions in the incidence of tooth decay in comparison to that in the control rats. some reduction in caries occurrence was observed. The effect of the amount of protein. No carious lesions were noted when mineralized whole milk diets were fed. VI. The number of cavities observed was comparable to that observed when the ration contained 20 parts of lard. fat and carbohydrate in the diet on the incidence and extent of carious lesions Schweigert BS. Effects of dietary composition on tooth decay in the albino rat Shaw JH. the type of caging arrangement employed throughout the experimental period was not found to influence tooth decay in the white rat. The addition of 10% of sucrose by weight to either mineralized fresh or evaporated milk did not result in any appreciable increase in tooth decay above the average for the animals on the comparable milk diets alone. In contrast. J Nutr 31: 439-447 (1946) Summary: The isocaloric substitution of 10 or 20 parts of lard for sucrose in a purified ration reduced the incidence and extent of carious lesions in the cotton rat in proportion to the amount of lard added. When the casein content of the diet was increased from 24 to 50% at the expense of sucrose. J Nutr 41:13-23 (1950) Summary: Weanling caries-susceptible albino rats were used as the experimental subjects in a series of investigations to determine whether the initiation and development of carious lesions would be influenced by dietary procedures already demonstrated to alter the incidence of tooth decay in caries-susceptible cotton rats. the protective effect was additive. Zepplin M and Elvehjem CA.

no mention of this calcium phosphate sequestering property has been found in the literature. however. the phosphopeptide is of practical use in preventing the formation of calcium phosphate precipitates in culture media containing relatively high concentrations of phosphate and calcium ion. 128. p.5. The result supports prior evidence that the cariogenicity of diets containing heat-processed skimmilk powders and deficient in lysine is due under some conditions to a critical deficiency of lysine. An autoclaved mixture of casein. has obvious implications in regards to an understanding of the role of casein in calcium and phosphate transport and assimilation. The finding of a casein fragment with the property of solubilizing calcium phosphate. or preventing its precipitation at relatively high pH’s. One of these fractions exhibits. by the edition of 11% of casein in place of cornstarch to this diet. to a remarkable degree. An inhibitory effect of a lysine supplement on caries was observed using purified diets containing zein as a source of protein.472 Introduction: By the use of ion-exchange columns. Science. Caries severity was significantly reduced. Mendel and Wakeman salts compared with unautoclaved casein was associated with an increased incidence of caries when the casein content of the diet was 13% but not when it was 24%. and Hubbell. 4. This striking caries difference was accompanied by only a slight difference in rate of growth.GC MI Paste – Abstract booklet – October 2007 3. the property of sequestering calcium phosphate in the pH range from 7 to 10. Calcium phosphate sequestering phosphopeptide from casein Reeves RE and Latour NG. The combined results of these studies suggest that the quantity of protein in the diet may be an important factor in the development of cariogenicity by experimental diets. 8 . Apart from its interest in this connection. Protein factors and experimental rat caries Bavetta LA and McClure FJ. Nutr 63: 107-117 (1957) Summary: A cariogenic diet containing a roller-process skimmilk powder remained highly cariogenic after supplementation with known vitamins and essential amino acids. A supplement of blood albumin proved as effective as L-lysine in the reduction of caries produced by a lysine-deficientskimmilk powder diet. a number of crude phosphopeptide fractions have been separated from pancreatic casein hydrolyzates. lactose. Although numerous investigations have been made on phosphopeptides derived from casein by various enzymatic hydrolyses. Vol. Diets containing 13% of casein developed a high incidence of severe caries but caries was very limited with 24% casein in the diet.

Storey E. milk contains factors which can contribute to the prevention of dental caries.3. 6.GC MI Paste – Abstract booklet – October 2007 5. The molars of the offspring of the protein-deficient females were significantly smaller than those of the controls. Many of the third molars of these rats had missing cusps. despite its content of potentially cariogenic lactose. Shelton. 1977) and anticariogenic (Shaw. The reason for this apparent contradiction is due largely to the inherent problems of studying the effect of one item of food on dental health. much circumstantial evidence is accumulating that. Shaw JH and Sweeney EA. Rats born to females fed a high-protein: low-sucrose diet had a tendency toward an increased susceptibility to a periodontal syndrome and were significantly less susceptible to dental caries than the offspring of females fed the control diet. 1979 Introduction: Milk contains all the essential nutrients required for the growth and development of bones and teeth. Effects of various sucrose-casein ratios in purified diets on the teeth and supporting structures of rats Holloway PJ. 1961 Purified diets of varying carbohydrate:casein ratios were fed to female rats of the Harvard caries-susceptible strain for a stabilizing period of 28 days and then throughout pregnancy and lactation. 34:175-179. Dairy Tech. Aust. is uncertain since it has been described as being both cariogenic (Gardner. but the syndrome did not progress as rapidly as among the offspring of females on the high-protein:low-sucrose diet. Norwood and Eisenson. Offspring of females fed a low-protein: high-sucrose diet grew more slowly and their molar teeth erupted later than rats born to females fed a control diet with a normal protein content. A review of the effect of milk on dental caries Reynolds EC. Arch Oral Biol Vol. The offspring of the deficient females had a significantly higher susceptibility to caries than the offspring of the control females. Whether milk is beneficial for teeth once they erupt however. This change in size was due to a decrease in the distance between the outer borders of the dentine rather than to a reduction in enamel thickness. Ensfield and Wollman. Stephan. There was also a slight tendency for a breakdown of the periodontal structures among the offspring of females fed the low-protein:high-sucrose diet. 1966). pp. 1977. J.185-200. 9 . 1959. Nevertheless. Berkowotz and Forrester.

beta-casein and kappa-casein) markedly reduced the rate of hydroxyapatite dissolution. The results lead to the hypothesis that the anticariogenic effect of milk is attributable to a direct chemical influence on the caries process in the oral environment of the rat. Arch Oral Biol Vol. p. 8.34 Suppl 2:S52-6 Summary: A chromatography column containing hydroxyapatite beads was used to study the effect of different proteins on the rate of hydroxyapatite dissolution. 26. nor with the animals having a significantly altered bacterial composition of their dental plaque. 445 to 451. expressed as the change in nmol calcium released per min per nmol of phosphoprotein bound to hydroxyapatite. The four phosphoproteins tested (phosvitin. The effect of the protein in reducing the hydroxyapatite dissolution rate has been attributed to protein binding to the surface of hydroxyapatite. 1981 Summary: Supplementation of a cariogenic diet with pasteurized bovine milk substantially reduced the incidence of dental caries in both male and female Sprague-Dawley rats. Riley PF. Three nonphosphorylated proteins had a relatively smaller effect. Calcif Tissue Int 1982. Effect of milk on caries incidence and bacterial composition of dental plaque in the rat Reynolds EC and Johnson IH.GC MI Paste – Abstract booklet – October 2007 7. The reduction in caries incidence was not associated with the consumption of less of the cariogenic diet and more milk. The reduction in dissolution rate. Reynolds EC. The results are consistent with the proposition that phosphoproteins have a regulatory role in mineralization processes and could provide a mechanism by which dietary and salivary phosphoproteins exert an anticariogenic effect. Storey E. alpha sl-casein. 10 .p. Phosphoprotein inhibition of hydroxyapatite dissolution. increased with increasing number of phosphoserine residues of the protein.

39(3):1285-90 The adherence of Streptococcus mutans PK1 to hydroxyapatite disks pretreated with various acidic and basic proteins in imidazole buffer was studied. as shown by zeta potential measurement. Within the basic protein group. Within the acidic protein group. Infect Immun 1983 Mar. the acidic residue percentage of the adsorbed protein was negatively correlated with the number of bacteria adhering. Wong A. These results suggest a topical anticariogenic action for dietary protein. These results indicate the involvement of short-range hydrophobic and ionic interactions in bacterial adherence to protein-coated hydroxyapatite. The finding that a 2 per cent solution of whey protein reduced the extent of fissure caries in animals consuming a solid diet containing 26 per cent whey protein suggests that the anticariogenic action is mediated by the protein being in solution. Adsorption of a basic protein onto an hydroxyapatite disk enhanced or had no effect on bacterial adherence. del Rio A. whereas adsorption of an acidic protein reduced adherence.29(11):927-33 Summary: Casein (bovine milk phosphoprotein) at 2 per cent (w/v) in drinking water reduced the extent of fissure and smooth-surface caries of male Sprague-Dawley rats consuming a solid cariogenic diet. whereas the nonpolar residue percentage was positively correlated with bacterial adherence. Whey protein (the non-phosphorylated protein group of bovine milk) also at 2 per cent (w/v) in the drinking water produced a smaller reduction and only of fissure caries. or in the amount or frequency of cariogenic diet consumed. the basic residue percentage was correlated with the number of cells adhering. The long-range effect of the acidic proteins in reducing the number of bacteria adhering to hydroxyapatite was related to protein adsorption causing an increase in surface net negative charge. Effect of casein and whey-protein solutions on caries experience and feeding patterns of the rat Reynolds EC. There was no significant difference in salivary-gland function (as determined by protein concentration).GC MI Paste – Abstract booklet – October 2007 9. 10. Arch Oral Biol 1984. Effect of adsorbed protein on hydroxyapatite zeta potential and Streptococcus mutans adherence Reynolds EC. The effect of adsorbed protein on bacterial adherence was of both short and long range. Basic protein produced a net positive surface charge which facilitated adherence. 11 .

protein. Hefferren JJ. Beck EX and Beck FM.mutans infection than did rats fed cheese containing no sucrose. but this increase was not significant. cheese-like substrate withour butterfat or casein and low in calcium and phosphate. with and without sucrose. mutans levels and enamel caries lesions were recorded.mutans implantation. Since cheddar cheese with sucrose did not significantly interfere with S. These results support previous reports of the cariostatic potential of cheese and suggest that a substantial portion of the protection may be related to textural influences and casein and/or calcium-phosphate content of the cheese. on dental caries and recovery of Streptococcus mutans in rats Rosen S. There was an increase in caries in rats fed cheeses with 20% sucrose. 14:384-389. An additional group was fed an agar gel with 25% lactalbumin and 25% soybean oil to assess the caries inhibition provided by a moist. Food consumption. Rats fed cheese or powdered diets containing sucrose had significantly higher frequency of recovery and higher levels of S.mutans levels were lower in the mozzarella and cheddar cheese groups than in the cream cheese group. 12.mutans. Clayton R. Min DB. alternate meals of processed cheese spread and mozzarella cheese substancially reduced caries incidence on buccal. Effect of cheese. calcium. The amount of butterfat. aging or carbohydrate in the test substrates had no apparent effect upon the amount of caries inhibition observed in this model. J Dent Res 63(6):894-896. S.GC MI Paste – Abstract booklet – October 2007 11. 1984 The objective of this study was to determinate the effect of aged and young cheddar cheese with and without added sucrose on dental caries and the associated recovery of implanted Streptococcus mutans. but not sulcal tooth surfaces. weight gains. the cariostatic mechanism is apparently unrelated to a direct antimicrobial effect on S. Cariostatic evaluation of cheeses with diverse physical and compositional characteristics Harper DS. Compared to the cheddar and gel control groups. 12 . Caries Res 20:123-130 (1986) The caries-inhibitory potential of four cheeses with different permutations of texture. Osborn JC. The cream cheese group had more buccal caries than the gel group. Very little caries was observed in rats consuming cheese without sucrose. plaque S. 1982]. [Caries Res. levels of butterfat. This study confirms the low cariogenic potential and possible cariostatic activity of cheddar cheese in rats. Harper DS. aging. There was significantly greater caries activity in rats fed standard diets containing 20% or 5% sucrose (SLS or MIT 305) than in rats fed cheeses containing 20% sucrose. Herper WJ. Test substrates were alternated with a cariogenic powdered diet and each fed 14 times daily for 28 days to albino rats superinfected with Streptococcus mutans. June. phosphate and lactose were evaluated in rat caries test using a controlled frequency feeding machine in a design similar to that of Edgar et al.

As such. mutans that can be associated with both its selection in stagnant areas and its cariogenicity. or feeding frequency. Rev. mutans occurs after tooth eruption. calcium. mutans suggests that treatment strategies which interfere with the colonization of S. and casein. if this colonization is delayed until the fissure depths are occupied by other bacteria. All three mineral concentrates significantly reduced buccal caries. possibly S. plaque acidity. and phosphate to modify the cariogenicity of a powdered diet containing 20% sucrose. Although the mechanisms responsible for protection are not completely identified. substances containing Ca and P may contribute to the protective potential by reducing demineralization and/or promoting remineralization of enamel. Vol. Microbiol. there is the possibility that decay will not occur or its occurrence will be greatly reduced. 1987 Jan. 353-380 These data provide convincing. Hefferren JJ. and two of the three reduced sulcal caries by from 10 to 30%. Modification of food cariogenicity in rats by mineral-rich concentrates from milk Harper DS. cheese. Role of Streptococcus mutans in human dental decay Loesche WJ. and if the fissures become colonized in their depths. Casein may reduce demineralization by forming a protective coat on the enamel surface. and lactobacilli are human odontopathogens. sobrinus appear to be important primarily in smooth-surface decay and. as such. can reduce the caries-causing potential of cariogenic substrates as measured in various animal. then decay may be inevitable. evidence that S. 50. this study evaluated the ability of three casein-free milk mineral concentrates with various levels of whey protein. as monitored by a computer-based infrared activity monitor. 14. J Dent Res. 1986. and in vitro systems. p. sobrinus.GC MI Paste – Abstract booklet – October 2007 13. The analysis further shows that casein-free milk mineral fractions can modify the cariogenicity of sucrose-containing foods in a rat model. Osborn JC. Analysis of these data indicates that there were no significant differences among groups for weight gain. 13 . Other aciduric species such as S. This understanding of the ecology of S. may be a cariogenic determinant when rampant decay occurs. However.66(1):42-5 Dairy products. including milk. total food consumption. By means of a rat model. Clayton R. albeit substantial. Colonization by S. dental caries is a diagnosable and treatable infection (209). mutans may have a profound effect on the incidence of dental decay in human populations (182). mutans. Aciduricity appears to be the most consistent attribute of S.

Burgess RC.0) per day prevented sub-surface enamel demineralization caused by six exposures of a 3% sucrose-3% glucose-salt solution per day over a ten-day period.01). The concentration of calcium was significantly higher in plaque from the experimental side (32. One side of each appliance (experimental) was dipped in a 25% water extract of the cheese for five min. thereby increasing plaque calcium phosphate and acid-buffering capacity by the phosphoseryl. p less than 0. The incorporation of casein and its breakdown in plaque did not produce a significant change in the amount or composition of plaque bacteria.0) prevented sub-surface enamel demineralization over a ten-day period as shown by microradiography and microhardness. and a tryptic digest of alpha s1-casein (TD-casein) were studied. while the other side (control) was dipped in de-ionized water. Jenkins GN. The left side of the appliance was exposed to various sugar and salt solutions.36 micrograms/mg. while the right side was exposed to sugar and casein solutions. the appliance was returned to the subject's mouth. Effects of water-soluble components of cheese on experimental caries in humans Silva MF.66(1):38-41 The effect of water-soluble components of extra-old Cheddar cheese on experimental caries was tested by means of the seven-day intraoral cariogenicity test (ICT). Immediately thereafter. glutamyl. Intact alpha s1-casein and tryptic peptides were shown immunochemically to be incorporated into the inter-enamel plaque. These results show that cheese has one or more water-soluble components which reduce experimental caries in human subjects. histidyl.44 micrograms/mg) as compared with the control side (19. These procedures were repeated six times per day.66(6):1120-7 The ability of bovine milk phosphoprotein (casein) to be incorporated into plaque. J Dent Res 1987 Jan. and two 60-second rinses with 10% sucrose were performed. or TD-casein solution (pH 7. Two bovine enamel blocks were placed in each buccal flange of the dental appliances of five volunteers. The cheese-extract dippings reduced the cariogenicity of the sucrose by an average of 55. the difference was not statistically significant. The prevention of sub-surface demineralization of bovine enamel and change in plaque composition by casein in an intra-oral model Reynolds EC.90 micrograms/mg) than on the control side (9. 14 . Sodium caseinate. and aspartyl residues and indirectly through catabolism by plaque bacteria.7% (p less than 0. Sodium caseinate at a level of 2% w/v in a 3% sucrose-3% glucose-salt solution (pH 7. however. the major fraction alpha s1-casein. J Dent Res 1987 Jun.GC MI Paste – Abstract booklet – October 2007 15. prevent enamel sub-surface demineralization. Sandham HJ. and affect bacterial composition was determined using a modified intra-oral caries model.61 micrograms/mg). The concentration of plaque phosphorus was higher on the experimental side (12. The ability of casein and tryptic peptides to prevent enamel demineralization was related to their incorporation into plaque.01). alpha s1-casein. as assessed by enamel microhardness. Two exposures of a 2% w/v sodium caseinate. Neither the mean resting pH nor the mean minimum pH in response to sucrose was significantly different between the experimental and control sides. Supragingival plaque was collected and impacted into the left and right inter-enamel spaces. 16. The intra-oral model consisted of a removable appliance containing a left and right pair of bovine enamel slabs placed to simulate an approximal area.

6% w/w casein. and kappa and the histones H1 and H3 were extensively hydrolyzed by the salivary-sediment bacteria. Populations of Streptococcus mutans did not differ among the groups. A significant difference in the frequency of eating and drinking periods of the animals from the four groups could not be demonstrated. salivary output. None of the other proteins extensively hydrolyzed by salivary sediment. The increase in ammonium ion concentration not attributed to arginine hydrolysis was correlated with the content of glutaminyl plus asparaginyl residues of the proteins. the caseins alpha s1. Of the proteins studied. Young DA. beta. One chocolate confection contained 5. Pre-hydrolysis of the arginine-rich histone H3 and poly-L-arginine with trypsin resulted in a marked increase in putrescine produced. the caries experience of the animals consuming milk chocolate and caramel was significantly lower than that of the animals eating sucrose. and ammonia were measured by use of a combined pH electrode.5% v/v) in a lactate-salt medium with an initial pH of 5. highperformance liquid chromatography. 15 . calcium or phosphate concentrations between the animals on the two chocolate diets. The levels of fat and sucrose were the same in both confectionery. Actinomyces viscosus was not detected at the end of the experiment in any of the groups. served as substrates for putrescine or cadaverine production. The effects of cheese snacks were particularly dramatic on root-surface caries. saliva was collected from the animals and caries was assessed.66(6):1116-9 Rats that had had their submandibular/sublingual glands removed surgically. indicating that the lower caries experience was not due to a less frequent sugar challenge. Animals consuming the casein-enriched chocolate either ad libitum or programme-fed had significantly (p<0. The effects of cheese snacks on caries in desalivated rats Krobicka A. Forty Sprague-Dawley male rats infected with streptomycin-resistant Streptococcus mutans consumed the chocolate diets either ad libitum or in a Konig/Hofer programmed feeder.68(2):124-9 Proteins of known composition and structural characteristics were incubated (1. Caries Res 1987. J Dent Res 1989 Feb. calcium and phosphate levels of all animals were normal. However. J Dent Res 1987 Jun. Black CL. or salivary protein. Reduction of chocolate’s cariogenicity by supplementation with sodium caseinate Reynolds EC.001) lower smooth surface and fissure caries scores than the animals consuming the normal chocolate. The caries reduction was attributed to a topical effect that was related to the difference in the composition of the confectionery. The frequency of eating and drinking by the animals was monitored using a time-lapse video recorder. Confectionery composition and rat caries Reynolds EC. The only amine detected was the polyamine putrescine arising from the catabolism of arginine following the hydrolysis of the arginine-rich histone H3. Protein dissimilation by human salivary-sediment bacteria Reynolds EC. mutans recovered from the swabs of the molar teeth of animals on both diets. These observations may be relevant for elderly humans who are most likely to develop root surface caries. 21:445-451 The cariogenicity of two chocolate confections was compared using a rat caries model. 19. Black CL.6% w/w casein while the other contained 16. 18. 20. streptococci or streptomycin-resistant S. developed fewer and less severe caries lesions on coronal and root surfaces when fed cheese snacks in addition to a cariogenic diet than when fed additional cariogenic snacks or no additional snacks. After 35 days. Incubation of salivary-sediment bacteria with the caseins and the histone H3 resulted in an increase in ammonium ion concentration and an associated decrease in hydrogen ion concentration. The work confirms an earlier report of a topical anticariogenic effect of soluble caseinate and shows that it is possible to reduce the cariogenicity of chocolate by caseinate supplementation. respectively.2 for two hr at 37 degrees C. The casein enrichment was achieved by replacing the 18% w/w non-fat milk solids used normally in the manufacture of milk chocolate with soluble sodium caseinate. suggesting that the salivarysediment proteolytic activity was not "trypsin-like". There was also no significant difference in the number of total organisms. or both of these components were responsible for the caries reduction. There was no significant difference between the caries experience of animals consuming fudge or sucrose. and glutamate dehydrogenase. amines. although containing arginyl and lysyl residues. milk chocolate. Hydrolysis of the proteins was monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Pearson S. Hydrogen ion. The difference in caries activity was not attributed to a difference in salivary function as salivary protein. and their parotid ducts tied. 21:538-545 Sucrose. Bowen WH. There was no significant difference in the mean final body weight. Riley PF. The hydrolysis of these proteins was attributed to their relative lack of tertiary (folded) structure. The levels of protein (casein) and fat of the test diets were inversely correlated with the animals’ caries experience suggesting that one.GC MI Paste – Abstract booklet – October 2007 17. The results of this study demonstrate that cheese exerts a protective effect against coronal and root-surface caries in rats with a severely limited salivary function. caramel or fudge were fed to rats infected with streptomycin-resistant Streptococcus mutans and receiving essential nutrients by gavage. Caries Res 1987.0 mg/mL) with re-suspended salivary sediment (2.

For this indicator. The three groups were exposed to the same basic preventive program. unpublished) and their anticariogenic potential is presently being investigated. increased the level of plaque calcium and inorganic phosphate and prevented subsurface enamel demineralization in a human intra-oral caries model. there was no difference between the two experimental groups. Results for the plaque index were in agreement with those of the DMF(S) increment and the net progression of decay. J Dent Res. compared with 6. Cariogenicity of a confection supplemented with sodium caseinate at a palatable level Reynolds EC. 1990 Nov. A 24-month clinical study of the incidence and progression of dental caries in relation to consumption of chewing gum containing xylitol in school preventive programs Kandelman D. Gagnon G. 1987).24 surfaces. They were divided into two experimental groups (15% and 65% xylitol chewing gum distributed three times a day at school) and one control group (without chewing gum).GC MI Paste – Abstract booklet – October 2007 21. Chewing xylitol gum had a beneficial effect on the caries process for all types of tooth surfaces.69(11):1771-5 The effect of chewing gum containing xylitol on the incidence and progression of dental caries was tested in a sample of 274 children. It could be possible therefore to supplement confectionery with the phosphopeptides at a level of one tenth that of caseinate (e. 16 . Caries Res 1989. Results for the two groups chewing gum were similar.0%. aged eight and nine years. The phosphopeptides can be easily purified from a tryptic digest of caseinate by selective precipitation (Reynolds. 1982).23:368-370 Summary: Recently Reynolds (1987) showed that tryptic peptides of caseinate were incorporated into plaque. These phosphopeptides are approximately 10% by weight of caseinate and are tasteless (Swaisgood. 2. Children who chewed gum had a significantly lower net progression of decay (progressionsreversals) over a 24-month period than did the controls.g.0%) without affecting palatability or consistency but still achieving the reduction in cariogenicity obtained with caseinate at 20. of low socio-economic status and high caries rate. It was concluded that the tryptic peptides associated with the anticariogenic effect were the calcium phosphate sequestering phosphopeptides of αs-caseinate and β-caseinate (Reynolds. The two experimental groups had a DMF(S) increment of 2. and especially for bucco-lingual surfaces.06 surfaces for the control group. Black CL. 22.

GC MI Paste – Abstract booklet – October 2007

23. Efficient solution-phase synthesis of multiple O-phosphoseryl-containing peptides related to casein and statherin
Perich JW, Kelly DP, Reynolds EC. Int J Pept Protein Res 1992 Aug;40(2):81-8 The multiple Ser(P)-containing peptides, H-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-NHMe.TFA, H-Asp-Ser(P)-Ser(P)-Glu-Glu-NHMe.TFA and H-GluSer(P)-Ser(P)-Glu-Glu-NHMe.TFA were prepared by the use of Boc-Ser(PO3Ph2)-OH in the Boc mode of solution phase peptide synthesis followed by platinum-mediated hydrogenolytic de-protection of the Ser(PO3Ph2)-containing peptides. The protected peptides were assembled using the mixed anhydride coupling methods with 40% TFA/CH2Cl2 used for removal of the Boc group from intermediate Bocprotected peptides.

24. The use of synthetic phosphopeptides for epitope mapping of the αS1-casein phosphopeptide segment 59-79
Perich JW and Reynolds EC. Bioorg Med Chem Lett, Vol.2, No.9, pp. 1153-1154, 1992 Through the use of synthetic Ser- and Ser(P)-containing peptides, both sequences –Ser(P)-Ser(P)-Ser(P)- and Ile-Val-Pro-Asn-Ser(P)-ValGlu-Glu of the tryptic phosphopeptide segment 59-79 of αS1-casein were shown to be recognizd by anti-αS1-casein polyclonal antibodies in a competitive ELISA. Since replacement of Ser(P) with Ser in the later peptide resulted in complete loss of antibody recognition, this indicates that the phosphorylated seryl residue is a critical residue for antibody recognition.

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GC MI Paste – Abstract booklet – October 2007

25. An in situ model for simultaneous assessment of inhibition of demineraliza-tion and enhancement of remineralization
Featherstone JD, Zero DT. J Dent Res 1992 Apr;71 Spec No:804-10 In situ models to assess the ability of oral care products or food components to enhance remineralization and/or inhibit demineralization of tooth enamel or roots must be very carefully designed to minimize the confounding effects of the many variables involved. Controlling these variables as closely as possible is essential if meaningful answers are to be obtained from the models. We have developed an in situ model which combines the experience of several groups. Detailed screening of subjects is essential. Selection criteria should include good general health, good dental health, mandibular partial denture, at least eight natural teeth, no active caries lesions, known fluoride history, normal salivary function, and no medications that affect salivary function. Each subject carries a sound enamel slab and an enamel slab with a preformed caries-like lesion (demineralized in vitro) in his/her denture on each side of the mouth for test periods of two or four weeks. The demineralization challenge is controlled by extra-oral immersion of the appliances in sucrose daily. Daily product exposure or daily food component exposure is used as desired. Compliance indicators and a diet diary are included. Whole saliva flow rate (unstimulated), plaque acidogenicity, and salivary fluoride are monitored during the test periods. At the end of the test period, the test slabs are assessed for mineral change, after being sectioned, by means of cross-sectional microhardness or microradiography. The mineral loss or gain (delta M, microns x vol%), compared with adjacent control sections retained in the lab, is calculated as change in delta Z (microns x vol%), namely, delta M = delta ZTEST - delta ZCONTROL.

26. The analysis of multiple phosphoseryl-containing casein peptides using capillary zone electrophoresis
Adamson N, Riley PF, Reynolds EC. J Chromatogr 1993 Sep 3;646(2):391-6 Multiple phosphoseryl-containing sequences of peptides and proteins stabilize amorphous calcium phosphate at neutral and alkaline pH and have been implicated in the nucleation/regulation of biomineralization. In an approach to analyze these peptides using capillary zone electrophoresis (CZE) we have attempted to relate the absolute electrophoretic mobility of various casein phosphopeptides to their physicochemical properties. Multiple phosphoseryl-containing peptides were selectively precipitated from enzymic digests of sodium caseinate and further purified using RP-HPLC and anion-exchange fast protein liquid chromatography. Purified fractions were then analyzed by CZE. Absolute electrophoretic mobilities of 13 peptides were determined by measurement of migration times relative to that of a neutral marker, mesityl oxide. A linear relationship (r2 = 0.993) was obtained between absolute electrophoretic mobility and q/M(r)2/3 where q is the net negative charge of the peptide calculated using relevant pKa values and M(r) is the molecular mass. M(r)2/3 is a measure of the surface area of a sphere that has a volume proportional to the M(r) of the peptide and relates to the frictional drag exerted on the peptide during electrophoretic migration. As absolute electrophoretic mobility is influenced by charge and size CZE can be used to monitor peptide phosphorylation, dephosphorylation, deamidation and truncation. This technique therefore would be suitable for quantitative analysis of peptide substrates in kinase and phosphatase studies. In conclusion CZE is a rapid and efficient technique for the resolution of multiple phosphoseryl-containing peptides from enzymic digests of casein.

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GC MI Paste – Abstract booklet – October 2007

27. A selective precipitation purification procedure for multiple phosphoseryl-containing peptides and methods for their identification
Reynolds EC, Riley PF, Adamson NJ. Anal Biochem 1994 Mar; 217(2):277-84 Multiple phosphoseryl-containing sequences of proteins stabilize amorphous calcium phosphate and have been implicated in the regulation of biomineralization, protein structure, and enzyme activity. To facilitate studies on the identification and characterization of multiple phosphoseryl-containing sequences of proteins we have developed a simple and efficient purification procedure involving precipitation of Ca2+/ethanol-induced aggregates of the multiple phosphoseryl-containing peptides from enzymic digests. The multiple phosphoserylcontaining peptides of a tryptic digest of casein were selectively precipitated using Ca2+ (20 mol/mol protein) and 50% (v/v) ethanol at pH 3.5, 4.6, and 8.0. The individual peptides of the precipitates were purified using anion-exchange fast-performance liquid chromatography and reversed-phase HPLC and then identified by solid-phase sequence analysis and amino acid composition analysis after vapor-phase hydrolysis. Prior to sequence analysis the phosphopeptides were covalently coupled to arylamine membranes and the phosphoseryl residues converted to S-ethylcysteinyl residues by calcium-ion-catalyzed beta-elimination in the presence of ethanethiol. The modified peptides were sequenced using an Applied Biosystems Inc. automated protein sequencer fitted with a membrane cartridge. Only peptides containing the cluster sequence -Ser(P)-Ser(P)-Ser(P)- were precipitated by Ca2+/ethanol at pH 3.5. The pH 4.6 precipitate contained all the cluster peptides plus two diphosphorylated peptides containing -Ser(P)-Glu-Ser(P)- and -Ser(P)-Thr-Ser(P)-. At pH 8.0, a monophosphorylated peptide containing -Ser(P)-Glu-Glu- was also present in the precipitate with the diphosphorylated and cluster peptides. The recoveries of the peptides in the pH 8.0 selective precipitate ranged from 83 to 95% of that present in the hydrolysate.

28. High performance capillary electrophoresis of casein phosphopeptides containing 2-5 phosphoseryl residues: Relationship between absolute electrophoretic mobility and peptide charge and size
Adamson NJ, Reynolds EC. Elecrophoresis 1995, 16, 525-528 Multiple phosphoseryl-containing peptides from casein containing the cluster sequence –Ser(P)-Ser(P)-Ser(P)-Glu-Glu- stabilize amorphous calcium phosphate at neutral and alkaline pH and have been shown to be anticariogenic in various in vitro, animal and human experiments. In an approach to obtain insight into the structure and function of these peptides, we previously developed a method for their analysis using / high-performance capillary electrophoresis (HPCE). A linear relationship was obtained between absolute electrophoretic mobility and q/M² ³. / 0.43 / Di-and triphosphorylated peptide absolute electrophoretic mobility correlated with both q/M² ³ and ln(q+l)/n . However, for both the q/M² ³ 0.43 and ln(q+l)/n relationships with absolute electrophoretic mobility, the di- and triphosphorylated peptides formed a separate linear relationship to that of the cluster peptides. From these relationships, a di- or triphosphorylated peptide exhibited a greater absolute / 0.43 electrophoretic mobility than a corresponding cluster peptide with the same q/M² ³ or ln(q+l)/n value. This implies that the cluster peptides have a reduced effective net negative charge due to an electrical double layer effect and/or a larger hydrodynamic volume or a more extended structure relative to the di- and triphosphorylated peptides which is associated with a greater frictional drag electrophoretic migration.

19

74(6):1272-9 Casein phosphopeptides (CPP) stabilize calcium phosphate through the formation of casein-phosphopeptide amorphous calcium-phosphate complexes (CPP-CP). 1. phosphoseryl-containing. Ac-GluSer(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-NHMe.0% w/v) were applied to the animals' molar teeth twice daily. The anticariogenic effects of CPP-CP and fluoride were additive.5.5% w/v). corresponding to the common sequence in the CPP. Reynolds EC.i + 1). confirming that this calcium-phosphate-stabilizing portion of the casein phospho-peptides is associated with anticariogenicity. Johnson IH. NH to C H coupling constants and the observation of slowly exchanging amide protons. Black CL. Riley PF. the non-phosphorylated peptides of a casein tryptic digest (0. The CPP-CP significantly reduced caries activity in a dose-response fashion.I α α +5) NOE connectivities. medium-range (I.0% CPP-CP producing 55% and 46% reductions in smooth surface and fissure caries activity. The secondary structure of the peptide was characterized by sequential (i.1. or the calcium-phosphate complex of a synthetic octapeptide.2.GC MI Paste – Abstract booklet – October 2007 29. tryptic peptide αs1-casein(59-79) containing the phosphorylated motif –SSSEE-. Perich JW. Cain CJ.5% CPP-CP plus 500 ppm F had significantly lower caries activity than those animals receiving either CPP-CP or fluoride alone. Other groups of animals received solutions containing 500 ppm F. since animals receiving 0. The animals consumed a defined cariogenic diet free of dairy products. with 1.I + 2/3/4). 20 . being similar to that of 500 ppm F. Spectra have been recorded in the presence of excess + Ca² and at three different values of sample pH to characterize the changes in peptide conformation as calcium binds to the phosphorylated residues. Anticariogenicity of calcium phosphate complexes of tryptic casein phosphopeptides in the rat Reynolds EC. respectively. The tryptic digest of casein with the phosphopeptides selectively removed showed no anticariogenic activity. A 1H-NMR study of the casein phosphopeptide αS1-casein (59-79) Huq NL. The synthetic octapeptidecalcium phosphate complex significantly reduced caries activity. The ability of CPP-CP to reduce caries activity was investigated by use of specific-pathogen-free rats inoculated with Streptococcus sobrinus. 0. Biochim Biophys Acta 1247 (1995) 201-208 Complete sequence-specific resonance assignments have been determined for a calcium phosphate sequestering. and residues E61 to ∑67 involved in a looptype structure. Cros KJ. Solutions (100 microL) of the CPP-CP (0. C H chemical shifts. The CPP-CP did not significantly affect the level of S. J Dent Res 1995 Jun. 30. 0. sobrinus in fissure plaque. and long-range (I. Webber FL. Two structured regions have been identified: residues P73 to V76 implicated in β-turn conformations.

and food additives for the prevention of dental caries.0 precipitate (13. In situ caries models Zero DT.0 precipitates. and methionine oxidation. The recovery of the ACPP from the original caseinate was similar for both the pH 4. a combination of hard tissue substrates--including sound.GC MI Paste – Abstract booklet – October 2007 31. In the pH 8. the single-step selective precipitation with Ca2+/ethanol should be performed at pH 4. Sodium caseinate (10% w/v) was hydrolyzed by Novo trypsin (commercial grade) at 50°C for 2 h and CPP were purified from the 2+ acid clarified hydrolysate by a single-step selective precipitation procedure involving Ca (20 mol/mol casein) and ethanol (50% v/v) at pH 4. The yield of the pH 8. mouthwash. ethical considerations. 32.85 ± 0. s1(59-79). sound analytical support. in order to prepare casein phosphopeptides predominantly containing the cluster sequence -Ser(P)-Ser(P)Ser(P)-Glu-Glu-. 21 . surface-softened lesions and subsurface lesions--may be necessary to model all aspects of caries progression and prevention successfully. were detected. ACPP can be produced under industrially relevant conditions with only minor modifications such as slight truncation. Each parameter must be carefully considered in relation to the objectives of the research. study design requirements.9 mol %) due to increased recoveries of the diphosphorylated peptides (24. discussion 231-4 By using in situ models. In conclusion.0 precipitate the cluster peptides represented a smaller proportion of the total peptides (61. EC Reynolds.6 rather than pH 8. the physical design of the model. and a knowledge of how to work with research subjects to achieve a high level of compliance. However.4 mol %) as well as the additional recovery of the monophosphorylated peptide (33-48) (13. we have the potential to study both fundamental aspects of the caries process as well as more applied research problems such as the effect of food on dental caries and the role of fluoride in caries prevention in human subjects without actually causing caries in the natural dentition. In an approach to develop a commercial-scale process for the production of ACPP we have comprehensively characterized casein phosphopeptides (CPP) produced under industrially relevant conditions.4 mol %) ACPP cluster peptides with small amounts of the diphosphorylated peptides (13.7 mol %) indicating increased cross-linking by Ca2+ at the higher pH.04 ± 0. The individual peptides of the CPP preparations were purified by reversed-phase high-performance liquid chromatography (HPLC) and then identified by amino acid composition and sequence analyses. the type of hard tissue substrate and the method of assessing mineral status.0.6 and 8. deamidation. s1(43-58) and s2(126-136). The key experimental parameters that need to be considered in the development of an in situ model are the characteristics of the subject panel. However.48 wt % of the caseinate) was slightly higher than that of the pH 4. resulting in minor truncation of some peptides.6 or 8. and the study design and clinical protocol. Characterization of tryptic casein phosphopeptides prepared under industriallyorelevantoconditions NJ Adamson.0. impact on clinical relevance. The major source of variation associated with in situ models should be of biological and not experimental origin. Also some deamidation and methionine oxidation of one peptide. The design and conduct of proper in situ model studies require a clear understanding of the caries process.and have commercial potential as toothpaste.6 mol %). Given the complex nature of caries. and impact on the control of variation.30 wt % of the caseinate).9(3):214-30. Slight chymotryptic activity was detected in the industrial-grade enzyme.6 precipitate (11. Biotec Bioeng 1995 45:196-204 Anticariogenic casein phosphopeptides (ACPP) contain the cluster sequence -Ser(P)-Ser(P)-Ser(P)-Glu-Glu. Internal validation of in situ models using fluoride dose-response controls is considered to be necessary for studies evaluating the efficacy of new fluoride dentifrice formulations. Adv Dent Res 1995 Nov.6 precipitate contained predominantly (86. the pH 4.

Evaluation strategies that focus on only one potential mode of action (e. mutans significantly. Wahlgren NM and Drakenberg T. may explain its modified cariogenic potential. can be added to sugar-containing foods and therefore have commercial potential as an anti-cariogenic additive to foods and toothpastes. Using laboratory. 22 . the caution that must be applied in extrapolating laboratory data to predict clinical performance is discussed. Golliard M. The proposed mechanism of anticariogenicity for the CPP-ACP is that they localize ACP in dental plaque which buffers the free calcium and phosphate ion activities thereby helping to maintain a state of supersaturation with respect to tooth enamel. but. Dairy products and dental health Reynolds EC. The casein phosphopeptides (CPP) are produced from a tryptic digest of the milk protein casein by aggregation with calcium phosphate and purification by ultrafiltration. Incorporation of caseinoglycomacropeptide and caseinophosphopeptide into the salivary pellicle inhibits adherence of mutans streptococci Schupbach P. and this paper reviews some of the pre-clinical models that have been utilized in their evaluation and some of the pitfalls that must be avoided. in a proportion of adsorbed conformational states.9(3):304-11. The CPP-ACP. unlike fluoride. All samples were prepared for electron microscopy by high-pressure freezing followed by freeze-substitution. A wide range of non-fluoride anti-caries agents has been postulated. 35. The resulting pellicle was washed and incubated with caseinoglycomacropeptide (CGMP) and/or caseinophosphopeptide (CPP) labeled with 17. Through their multiple phosphoseryl residues the CPP bind to clusters of amorphous calcium phosphate (ACP) in metastable solution. Except for some reports associating nursing bottle caries with milk consumption. Although laboratory and in situ data predict anti-caries efficacy for sodium trimetaphosphate in combination with fluoride. the search for more effective alternative therapies continues.GC MI Paste – Abstract booklet – October 2007 33. is discussed. the lack of suitable calibration methods. A possible explanation for these findings is that milk components are incorporated into the salivary pellicle. inhibition of demineralization) may overestimate the true clinical efficacy which may arise from a combination of two or more effects (e. Finally. Adv Dent Res. This depresses demineralization and enhances remineralisation.. inhibition of demineralization and stimulation of remineralization). Ability of a β-casein phosphopeptide to modulate the precipitation of calcium phosphate by forming amorphous dicalcium phosphate nano-clusters Holt C. the disease remains a major public health problem. The ability of casein to form nanoclusters in milk suggests a more general mechanism for avoiding pathological calcification and regulating calcium flow in tissues and biological fluids exposed to or containing high concentrations of calcium. The nanoclusters were prepared from an undersaturated solution of salts and the peptide by raising the pH homogeneously from about 5. sobrinus and S. thereby reducing the adherence of S. The peptide is linked to the calcium phosphate through its sequence of phosphorylated residues. A reduction in the Streptococcus sobrinus population in the oral microbiota of animals fed diets supplemented with these milk components was consistently observed. 36. some comments on the appropriateness of laboratory evaluation strategiesoareomade. together with the increased remineralization potential of this biofilm. discussion 312-4 While fluoride is an effective anti-caries agent.g. J (1996) 314 (1035–1039) The ability of casein in the form of colloidal-sized casein micelles to modulate the phase separation of calcium phosphate during milk secretion is adapted to produce nanometre-sized particles of calcium phosphate stabilized by a casein phosphopeptide (nanoclusters). This hypothesis was tested in vitro by the incubation of bovine enamel discs with unstimulated saliva.g. 1779-1788 (1996) The protective effects of milk and milk products against dental caries have been demonstrated in many animal studies. This protein was identified with a mouse anti-human serum albumin followed by goat anti-mouse IgG labeled with 25-nm gold particles. Chemical analysis and IR spectroscopy showed that they comprise an amorphous dicalcium phosphate bound to the phosphopeptide. Biochem. We have shown that this effect was mediated by micellar casein or caseinopeptide derivatives. this was not found in three-year clinical trials. Vol 75. 1995 Nov. as well as by transmission electron microscopy. Rouvet M and Guggenheim B. The CPP have a remarkable ability to stabilise calcium phosphate in solution and substantially increase the level of calcium phosphate in dental plaque. Even though in most developed countries the prevalence of dental caries has decreased through the use of fluorides. Multinuclear NMR spectroscopy of the cluster solutions showed that the small ions and free peptide in the solution were in a state of dynamic exchange with the nanoclusters. It was demonstrated by high-resolution scanning electron microscopy with back-scattered electron imaging. Neeser JR. preventing their growth to the critical size required for nucleation and precipitation. 34. the termini retain the conformational freedom of the unbound peptide. Proceedings of the Nutrition Society of Australia (1995) 19 Dental caries (tooth decay) is initiated via the demineralization of tooth hard tissue by organic acids from the fermentation of dietary sugar by dental plaque odontopathogenic bacteria. dairy products have been recognized for over 40 years as exhibiting an anticaries effect. A possible reason for this. J Dent Res. The resulting ecological shifts. sobrinus. Using data on the potential anti-caries efficacy of phosphopeptides obtained from casein.and 12-nm gold particles. confirming previous findings. that both peptides were incorporated into the pellicle in exchange for albumin. Incorporation of CGMP and/or CPP into salivary pellicles reduced the adherence of both S.5 to 6. It is suggested that the calcium and phosphate-rich micellar casein or caseinopeptides are incorporated into the pellicle. animal and human in situ caries models it has been shown that casein phosphopeptide amorphous calcium phosphate complexes (CPP-ACP) exhibit an anticariogenic activity.. Role of models in assessing new agents for caries prevention-non-fluorideosystems Roberts AJ.7 with urea plus urease.

No 5.F. G. 38. J Dent Res 1997 Sep.have been demonstrated to significantly reduce the level of dental caries (tooth decay) in animal and human experiments and also to repair early stages of decay. P. F.9 +/. localize ACP in dental plaque. Morgan. Advances in enamel remineralization: anticariogenic casein phosphopeptide amorphous calcium phosphate E. N. Webber. The CPP-ACP. in particular the neutral ion pair CaHPO4(0). maintain high-concentration gradients of calcium and phosphate ions and ion pairs into the subsurface lesion and thus effect high rates of enameloremineralization.2H2O). The CPP localize ACP at the tooth surface providing a reservoir of calcium and phosphate ions thereby helping to maintain a state of supersaturation with respect to tooth enamel. The casein phosphopeptides (CPP) are produced from a tryptic digest of the milk protein casein by aggregation with calcium phosphate and purification by ultrafiltration. by stabilizing calcium phosphate in solution. which decreased the concentrations of free calcium and phosphate ions and increased the level of CPP-bound ACP. the solutions were stabilized by the CPP such that spontaneous precipitation of calcium phosphate did not occur. Through their multiple phosphoseryl residues. After a ten-day remineralization period. Cross.0% CPPcalcium phosphate (pH 7. Remineralization was not significantly correlated with either the CPP-bound ACP of the degrees of saturation for hydroxyapatite. F. P. 18(1):8-16 Using laboratory. unlike fluoride. Eakins.9 +/. The CPP. M. preventing their growth to the critical size required for nucleation and precipitation. pp. Vol. 40. enamel lesions were sectioned.V. The CPP have a remarkable ability to stabilize calcium phosphate in solution and substantially increase the level of calcium phosphate in dental plaque. with the 1. Spec Care Dentist 1998 Jan-Feb. The proposed mechanism of anticariogenicity for the CPP-ACP is that they localize ACP in dental plaque.iAnticariogenicocaseinophosphopeptides Reynolds EC. Cai. Conformational and binding studies have shown that all the phosphoseryl residues are important in interaction with ACP. animal and human in situ caries models. octacalcium phosphate.J. 23 . J Clin Dent 10: 86-88. preventing their growth to the critical size required for nucleation and phase transformations. 1999 Proteins and peptides containing clusters of phosphoseryl residues have been shown to stabilize amorphous calcium phosphate in solution. animal.8 x 10(-8) mol hydroxyapatite/m2/s. Remineralization of enamel subsurface lesions by casein phosphopeptide-stabilized calcium phosphate solutions Reynolds EC. can be added to sugar-containing foods as well as oral care products for the control of dental caries.0) solution replacing 63. or ACP. subjected to microradiography. 295-303. C. 39.1% of mineral lost at an averaged rate of 3. preventing their growth to the critical size required for nucleation and phase transformation. investigators have shown that casein phosphopeptide amorphous calcium phosphate complexes (CPP-ACP) exhibit an anticariogenic activity. Although most of the remineralizing solutions were supersaturated with respect to the amorphous and crystalline calcium phosphate phases. Talbo. All solutions deposited mineral into the bodies of the lesions.GC MI Paste – Abstract booklet – October 2007 37. and are anticariogenic in animal and in situ human caries model. CPP-stabilized calcium phosphate solutions were shown to remineralize subsurface lesions in human third-molar enamel. The remineralizing capacity was greater for the solutions with the higher levels of CPP-stabilized free calcium and phosphate ions. D. Huq. and the mineral content determined by microdensitometry. unlike fluoride. K. Through their multiple phosphoseryl residues the peptides bind to forming nanoclusters of amorphous calcium phosphate (ACP) in metastable solution. Black. thereby helping to maintain a state of supersaturation with respect to tooth enamel depressing demineralization and enhancing remineralization. Shen.C. Casein phosphopeptides (CPP) containing the cluster sequence –Ser(P)-Ser(P)-Ser(P)-Glu-Glu. Solutions were used to examine the effect of CPP-calcium phosphate concentration on remineralization. J.0. but his was attributed to the significant correlation of remineralization with the activity gradients from the solution into the lesion of some calcium phosphate ions and ion pairs. Through their multiple phosphoseryl residues the CPP bind to forming nanoclusters of ACP in metastable solution.76(9):1587-95 Casein phosphopeptides (CPP) stabilize amorphous calcium phosphate (ACP).L. These peptides have a remarkable ability to stabilize calcium phosphate in solution as amorphous calcium phosphate (ACP). 1999 Casein phosphopeptides (CPP) are multiphosphorylated peptides from an enzymatic digest of the bovine milk protein casein.L. The CPP-ACP have potential as an additive to foods and oral care products for the control of dental caries. the CPP bind to forming clusters of amorphous calcium phosphate (ACP) in metastable solution. Reynolds. remineralization was significantly correlated with the degree of saturation for dicalcium phosphate dihydrate (CaHPO4. which buffers the free calcium and phosphate lon activities. Perich. Anticariogenic complexes of amorphous calcium phosphate stabilized by casein phosphopeptides: a review Reynolds EC. Riley. The casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) nanoclusters have been shown to localize at the tooth surface and prevent caries in laboratory. Other solutions were used to examine the effect of increasing pH.W. 6. thereby helping to maintain a state of supersaturation with respect to tooth enamel depressing demineralization and enhancing remineralization. In this vitro study. and have been implicated in the regulation of biomineralisation processes. Prot Peptide Lett. The CPP-ACP. The proposed anticariogenic mechanism for CPP-ACP is the localization of ACP at the tooth surface which buffers the free calcium and phosphate ion activities.20. and human in situ caries models. can be added to sugar-containing foods and therefore have commercial potential as an additive to foods as well as to toothpastes and mouthwashes for the control of dental caries. However. The CPP-ACP have also been shown to remineralize enamel subsurface lesions in vitro and in situ when delivered in a sugar-free chewing gum.

The progress of the caries lesion is decelerated and remineralization accelerated. phosphate/acid phosphate.and CO3-poor dental apatites which are more resistant to subsequent acid challenges. FHA. The caries process is initiated by the dissolution of the tooth mineral (calcium carbonatehydroxyapatite) by organic acids (lactic and acetic acid) produced by plaque bacteria acting on dietary carbohydrates or by lowered pH from ingested food and drink. forms directly or indirectly by transformation of the other calcium phosphates. thereby helping to maintain a state of supersaturation with respect to tooth enamel depressing demineralization and enhancing remineralization. leading to the formations and transformations of different types of calcium phosphates.GC MI Paste – Abstract booklet – October 2007 41. a development which will add significantly to dentists’ armoury of tools in preventive dentistry.. The emineralization/remineralization processes consist of the dissolution of F-poor. J Clin Dent 10: 65-73. is most effective in inhibiting the dissolution of synthetic or enamel apatite.g. The role of phosphopeptides in caries prevention Reynolds EC. The dissolution increases the concentration of calcium. CaF2) associated with the demineralization/remineralization processes. The CPP-ACP therefore have exciting potential for use in oral care products and foods to help prevent dental caries and to help repair early stages of the disease. 1999 Enamel and dentin caries occurs as a result of a shift in the equilibrium between demineralization and remineralization processes. This paper reviews the calcium phosphates and other calcium compounds (e. (Ca+F) or (P+F) or F alone. Mg. Dental Perspectives 1999 3:6-7 Conclusion: The anticariogenicity of the CPP-ACP has been demonstrated in the rat and human in situ caries models.and CO3-rich dental apatites and the precipitation of F-rich. Caries lesion can be arrested by timely remineralization strategies.in solution. In the presence of low levels of fluoride (F-) ions. carbonate/bicarbonate ions in the microenvironment of the caries lesion. with the demineralization process predominating. 24 . at higher concentrations. phosphate (P) and fluoride (F) ions in solution compared to the combined presence of (Ca+P). Calcium phosphates in demineralization/remineralization processes Zapanta LeGeros R. CaF2-like materials form which can dissolve to form FHA and provide a supply of F. The CPP-ACP has also been shown to remineralize enamel subsurface lesions in vitro and in situ when delivered in a sugar-free gum. in F-containing enamel. Mg. (F. The simultaneous presence of calcium (Ca).OH)-apatite. 42. magnesium. calcium fluoride. The proposed anticariogenic mechanism for CPP-ACP is the localization of ACP at the tooth surface which buffers the free calcium and phosphate ion activities.

The use of casein phosphopeptides in oral care products for the prevention and treatment of early enamel caries Reynolds EC. we have determined the relative binding energies of the motif –Ser(P)-Ser(P)-Ser(P)-Glu-Glu to different crystalline surfaced of HA.stabilize amorphous calcium phosphate (ACP) in body fluids and bind with high affinity to crystalline calcium phosphate phases such as hydroxyapatite (HA) regulating crystal growth. Cross KJ and Reynolds EC. Binding of this motif to hydroxyapatite surfaces was investigated in this study using molecular modeling techniques. Using a threestep computational procedure.Mol. 2000. 44. J. These preferences are principally governed by electrostatic interactions between the crystal lattice and the peptide with the most stable conformers adopting structures where alternate residues exhibit backbone angles characteristic of a ß-strand and values of an α-helix or a distorted α-helix.Model. The results of this study are consistent with experimentally-derived data on the interaction of multiphosphorylated proteins/peptides with HA and have implications for the role of these proteins/peptides in calcium phosphate stabilisation and biomineralisation processes.GC MI Paste – Abstract booklet – October 2007 43. This analysis revealed preferences of the motif for (100) and (010) surfaces of the crystal and preferences for particular orientations on a given surface. allowing maximal interaction between the acidic side groups and surface calciums. 6. Aust J Dairy Technol 2000 55:1-6 (not published) 25 . Molecular modeling of a multiphosphorylated sequence motif bound to hydroxyapatite surfaces Huq NL. 35-47 Proteins and peptides containing the multiphosphorylated motif –Ser(P)-Ser(P)-Ser(P)-Glu-Glu.

lightness).ct. Subsequently.34(5):427-31 Casein phosphopeptides (CPP) stabilize amorphous calcium phosphate (ACP) and may be used to localize ACP in dental plaque. once in place.9 (Den Besten PK. Remineralisation of fluorotic enamel lesions by casein phosphopeptide .6 +/. The aim of this paper is to investigate these effects by measuring the affinity and capacity of Streptococcus mutans for CPP-ACP.2.0 +/. Giambro N.amorphous calcium fluorophosphate (CPP-ACFP) solution Roberts MJ. The relative whiteness of the lesions before and after remineralisation was quantitated by light reflectance (L value. 27:71-7). The percentage of mineral deposited increased with surface conditioning (80. with and without surface conditioning (5. providing a large calcium reservoir. Histological studies indicate fluorotic lesions are similar to enamel carious lesions. The results demonstrate that CPP-ACP binds with about twice the affinity of the bacterial cells for calcium up to a value of 0.25% w/v NaOCl for 20 minutes) prior to remineralisation with CPP-ACFP (5% w/v. Reynolds EC. premolar teeth (n=7) with fluorosis (TF=3) were sectioned into buccal and lingual samples and randomly assigned to two groups. 26 . The University of Melbourne).76:1587-1595].9.05).8%). and the remineralisation determined by microdensitometry.7% mineral within the fluorosed lesions. CPP-ACP will form a source of readily available calcium to inhibit demineralization. CPP-ACFP replaced 44. Binding characteristics of Streptococcus mutans for calcium and casein phosphopeptide Rose RK. Surface conditioning improved the effectiveness of the treatment with CPP-ACFP. p>0.4. 16 g/g wet weight cells.GC MI Paste – Abstract booklet – October 2007 45. Using the equilibrium dialysis system described by Rose and Hogg [Biochim Biophys Acta 1995. Abstract 54.6 +/.0 +/. 17:340-5).0 +/. 2000 Epidemiological studies show an increasing prevalence and severity of fluorosis amongst children in Australia especially for ThylstrupFejersfov (TF) indices of 2 and 3 (Riordan PJ.3.0 +/. subjected to microradiography.2. Caries Res 2000 Sep. maintaining a state of supersaturation with respect to tooth enamel. reducing demineralization and enhancing remineralization [Reynolds. These data can then be used to derive a true dissociation constant for CPP-ACP itself. Hence.0 to 72. J Dent Res 1997.05). CPP-ACP binds well to plaque.7 to 76. We conclude that treatment with CPP-ACFP significantly reduced the whiteness and partially remineralised fluorotic lesions.5. ANZ division. which is likely to restrict mineral loss during a cariogenic episode and provide a potential source of calcium for subsequent remineralization. CPP-ACFP treatment significantly reduced whiteness of fluorotic lesions (from 81. Literature-based values for non-fluorosed enamel are 69. 46.3. The reduction in whiteness increased with surface conditioning (from 85. To determinate whether CPP-ACFP could remineralise fluorotic lesions and improve the appearance. (Study supported by Research Committee School of Dental Science.1245:94-98]. p<0. pH 7) for 10 days. CPP-ACP will restrict the caries process.9.3 +/. Caries Res 1993. IADR. 76:1587-95).7. Overall. J Dent Res 1997. Fluorotic lesions were sectioned. In situ and in vitro studies have shown CPP-ACFP able to remineralise enamel carious sub-surface lesions (Reynolds EC. assessment of calcium binding by a plaque streptococcus at a fixed CPP-ACP concentration gives a series of CPP-ACP-influenced dissociation constants for calcium. Messer LB. Ped Dent 1995. Application of CPP-ACP to plaque may cause a transient rise in plaque fluid free calcium which may assist remineralization.

once in place. with calcium and phosphorus. providing a large calcium reservoir within the plaque and slowing diffusion of free calcium. CPP–ACP will restrict the caries process. 27 .3(1):27-30 Objective: To explore a new method for effective and safe prevention and treatment of caries. Zongtong Li. had a high affinity for hydroxyapatite and could enhance tooth remineralization. The aim here was to investigate these effects by measuring the effect of CPP–ACP on calcium diffusion in plaque.45(7):569-75 Casein phosphopeptides (CPP) stabilize amorphous calcium phosphate (ACP) and may be used to localize ACP in dental plaque. reducing demineralization and enhancing remineralization. Experimental study of phosphopeptide in promoting tooth remineralization Li Zhang. Using Dibdin's effusion system. Yingjie Dong. Effects of an anticariogenic casein phosphopeptide on calcium diffusion in streptococcal model dental plaques Rose RK. Conclusion: Phosphopeptide was effective in the prevention and treatment of tooth decay. ChinJ Dent Res 2000 May. 48. it could be safely applied. This demonstrated that by providing a large number of possible binding sites for calcium. maintaining a state of supersaturation with respect to tooth enamel. we searched for the mechanism and clinical application of tooth remineralization with phosphopeptide.1% CPP–ACP reduces the calcium diffusion coefficient by about 65% at pH 7 and 35% at pH 5. Results: Phosphopeptide. Methods: Following the theory of tooth remineralization. CPP–ACP binds well to plaque. Clinically. Arch Oral Biol 2000 Jul. Overall. calcium diffusion was measured in streptococcal model plaques. Hence. This is likely to restrict mineral loss during a cariogenic episode and provide a potential source of calcium for subsequent remineralization. 0.GC MI Paste – Abstract booklet – October 2007 47.

The studies involved a dose response of the CPP-ACP in sorbitol. sectioned and subjected to microradiography and densitometric image analysis.Abstract 0490 Tryptic phosphopeptides from milk caseins (CN). respectively. Supported by the National Health and Medical Research Council of Australia (Grant No. NL Huq. Supported by Warner-Lambert. 991486). human-enamel. Inc. we have been studying the structure-function relationships of the predominant peptides αsi-CN (59-79). The intra-oral appliances were inserted immediately before gum chewing for 20 min and were retained for a further 20 min immediately after gum chewing. half-slabs inset containing subsurface demineralized lesions. Each treatment was for 14 days duration and at the completion of each treatment the enamel slabs were removed. Thirty subjects in randomized. β-CN(1-25) adopts a preferred conformation when complexed with ACP. 28 . Powder diffraction X-ray crystallography confirmed that the calcium phosphate phase stabilized by β-CN (1-25) was non-crystalline. relative to the control gum. This was performed four times per day. complexed with amorphous calcium phosphate (ACP) exhibit anticariogenic activity based on laboratory. Structural studies of the β-casein phosphopeptide bound to amorphous calcium phosphate KJ Cross. The other half of each enamel slab was used as the control demineralized lesion. animal and human in situ caries models. 50.and xylitol-based sugar-free gum and also included a nil-treatment (no gum chewing) control. double-blind studies wore removable palatal appliances with six. General session. D Eakins. The use of either the sorbitol. Chiba 2001 .Abstract 0489 The ability of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) in sugar-free chewing gum to remineralize enamel subsurface lesions in a human in situ model was studied. 101% and 151%. cross-over. To understand the mechanisms of the anticariogenicity of the CPP-ACP complex. β-CN (1-25). and this conformation has similar features to that observed in β-CN(1-25) in the presence of calcium ions. paired with their respective demineralized control.or xylitol-based gum resulted in a dose-related increase in enamel remineralization with 0. a division of Pfizer.GC MI Paste – Abstract booklet – October 2007 49. 62%. EC Reynolds. A Nowicki. P Shen. Furthermore. IADR. IADR. Addition of CPP-ACP to either the sorbitol.4 mg of CPP-ACP producing an increase in enamel remineralization of 9%. In conclusion.8 mg and 56.19 mg. Enamel remineralization by chewing gum containing casein phosphopeptide-amorphous calcium phosphate EC Reynolds. nOe transfer from water solvent to solute revealed that specific protons are solvent exposed in the β-CN(1-25)-ACP. General session. The purpose of this study was to examine the solution structure of β-CN (1-25) complexed with calcium phosphate using Nuclear Magnetic Resonance (NMR) Spectroscopy. Chiba 2001 . 10 mg.60% enamel remineralization.05%. The nil treatment resulted in 3. 18. bound to calcium phosphate in solution and bound to hydroxyapatite crystals. J Vincent. embedded. The translational diffusion experiments indicate that the β-CN(1-25)-ACP particled observed by NMR are less than 10 nm in diameter. F Cai.or xylitol-based gum more than doubled the amount of remineralization with the average level of enamel remineralization for both gum types being 9. when bound to calcium ions.

in the presence of excess ammonium ions. Bicknell W. The secondary structure of the peptide was characterized by sequential and medium-range (i. 52. Ser(P)(17) to Glu(20) and Glu(21) to Thr(24) were implicated in beta-turn conformations. The aim of this study was to examine the structures of these complexes using a range of physico-chemical techniques. i+2/3/4. At low concentrations (1-10%. General session. The colloidal solutions of CPP-ACP exhibited a complex rheological behaviour. 29 . Comparison of the patterns of medium-range nOe connectivities in beta-casein-(1-25) with those in alpha(S1)-casein-(59-79) suggest that the two peptides have distinctly different conformations in the presence of calcium ions. Powder diffraction patterns have confirmed the amorphous nature of these complexes. powder diffraction X-ray crystallography. sodium ions and calcium ions. Huq NL. in separate experiments. Four structured regions were identified in the calcium-bound peptide: residues Arg(1) to Glu(4) were involved in a loop-type structure. D Eakins. 991486). Cation-dependent structural features of beta-casein-(1-25) Cross KJ. w/v) they behaved as free-flowing solutions whereas at higher concentrations (20-40% w/v) they transformed into thixotropic gels. including NMR spectroscopy. Biochem J 2001 May 15. Medium-range nuclear Overhauser effect (nOe) enhancements. Reynolds EC.356(Pt 1):277-86 Complete sequence-specific. EC Reynolds. which denotes an interaction between residue i and residue i+2. proton-resonance assignments have been determined for the calcium phosphate-stabilizing tryptic peptide beta-casein-(1-25) containing the phosphorylated sequence motif Ser(P)(17)-Ser(P)-Ser(P)-Glu-Glu(21).GC MI Paste – Abstract booklet – October 2007 51. IADR. Microfiltration studies suggest that the complexes can reach sizes over 0. We observed significant changes to chemical shifts for backbone and side-chain resonances that were dependent upon the nature of the cation present.Abstract 0491 Tryptic phosphopeptides derived from milk caseins are known to associate with amorphous calcium phosphate (ACP) forming stable complexes that behave as calcium phosphate delivery vehicles and are effective in the remineralization of early enamel lesions. and of the dephosphorylated peptide in the presence of excess sodium ions. NL Huq. Chiba 2001 . and residues Val(8) to Glu(11). despite having a high degree of sequential and functional similarity.2µm. and light scattering. Spectra of the peptide have been recorded. Ultrastructural studies of the casein phosphopeptide-amorphous calcium phosphate nanoclusters KJ Cross. Supported by the National Health and Medical Research Council of Australia (Grant No. and Halpha chemical shifts. were observed and also found to be cation dependent. i+3 or i+4 in the peptide) nOe connectivities. characteristic of small structured regions in the peptide. In conclusion. NMR studies suggest that the inner core of these complexes are less than 10 nm. the CPP-ACP complexes consist of small. nano-sized sub-unit particles that are cross-linked to form much larger particles (>200nm) or a gel network.

MALDI-PSD-MS analysis of the phosphorylation sites of caseinomacro-peptide Talbo GH. This analysis showed that CMP is fully phosphorylated at Ser(149) and only partially phosphorylated at Ser(127. The aim of this study was to use matrix assisted laser desorption/ionization post source decay mass spectrometry (MALDI-PSD-MS) to identify the phosphorylation sites in the CMP sequence. 30 .GC MI Paste – Abstract booklet – October 2007 53.) Dehydroalanyl residues corresponding to the phosphoserines of CMP were detected upon MALDI-PSD-MS analysis suggesting that the phosphoryl bond in phosphoserine is very labile during PSD analysis such that the phosphoryl group may be lost before backbone fragmentation. Suckau D. CMP was isolated from a chymosin digest of casein by HPLC and then digested with endoproteinase Glu-C to generate peptides suitable for MALDI-PSD-MS analysis. The aglycosylated. phosphorylated form of CMP has been shown to exhibit antibacterial activity. 2001 Jul. CMP naturally exists in several forms due to extensive posttranslational modifications including glycosylation and phosphorylation. Encyclopedia of Dairy Sciences 2001. 54. Reynolds EC. Health aspects of dairy products – Dairy products in relation to caries prevention and oral health.22(7):1093-8 Caseinomacropeptide (CMP) is a 64 amino acid polypeptide corresponding to kappa-casein 106-169. Reynolds EC. Malkoski M. Peptides.

The CPP-ACP have potential as an additive to foods and oral care products for the control of dental caries. for measurement of the level of remineralization. relative to the control gum. At the completion of each treatment. This was performed four times per day for 14 days. animal and human in situ caries models. preventing their growth to the critical size required for nucleation and phase transformations. Nowicki A. and human in situ experiments. with 0. Vincent J. J Dent Res 2001 Dec. double-blind studies wore removable palatal appliances with six human-enamel half-slabs inset containing sub-surface demineralized lesions.0. These peptides have a remarkable ability to stabilize calcium phosphate in solution as amorphous calcium phosphate (ACP). 31 . Remineralization of early enamel caries by anticariogenic casein phosphopeptide – amorphous calcium phosphate nanocomplexes Reynolds EC. 102. Thirty subjects in randomized.Reynolds EC.GC MI Paste – Abstract booklet – October 2007 55.19. Cai F. embedded. The proposed anticariogenic mechanism for CPP-ACP is the localization of ACP at the tooth surface which buffers the free calcium and phosphate ion activities. respectively. animal. The CPP-ACP have also been shown to remineralize enamel subsurface lesions in situ when delivered in a sugar-free chewing gum. independent of gum weight or type.4 mg of CPP-ACP producing an increase in enamel remineralization of 9. The addition of CPPACP to either sorbitol. thereby helping to maintain a state of supersaturation with respect to tooth enamel depressing demineralization and enhancing remineralization. Dental Practice 2001 November / December Casein phosphopeptides (CPP) are multi-phosphorylated peptides from an enzymatic digest of the bovine milk protein casein. Through their multiple phosphoseryl residues the CPP bind to forming nanoclusters of ACP in metastable solution. and 56. Remineralization of enamel subsurface lesions by sugar-free chewing gum containing casein phosphopeptide -amorphous calcium phosphate Shen P. The appliances were inserted immediately before gum-chewing for 20 min and then retained for another 20 min. The aim of this study was to determine the ability of CPP-ACP in sugar-free chewing gum to remineralize enamel subsurface lesions in a human in situ model. 63. sectioned. and subjected to microradiography and densitometric image analysis. cross-over. 56. 10.8.or xylitol-based gum resulted in a dose-related increase in enamel remineralization. 18.80(12):2066-70 Casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP) exhibit anticariogenic potential in laboratory. The casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) nanocomplexes have been shown to localize at the tooth surface and prevent caries in laboratory. and 152%. the enamel half-slabs were paired with their respective demineralized control half-slabs.

25%) in vitro.25µm (SD 0.47. Results: The pH values of test solutions were higher than that of P (2. under constant agitation in a water bath.09%. 3.stabilised amorphous calcium phosphate (CPP-ACP) required to be added to Powerade™ to eliminate erosion of human enamel in vitro.17).27. 3.83.An in vitro investigation of the effects of casein phosphopeptide-stabilized amorphous calcium phosphate (CPP-ACP) on erosion of human dental enamel by a sports drink Ramalingam L.09% CPP-ACP: 0. Superficial surface irregularities were displayed for P+0. Type 1 etch patterns predominated with scattered areas of Type 3 in P alone and P+0. 0. Methods: Surface changes of 30 human enamel specimens were examined following immersion in Powerade™ (P) containing 0. P+0. P and double deionized water (DDW) were positive and negative controls respectively.125% and 0.43µm (SD 0.125% CPP-ACP: 0. General Session.063% CPP-ACP displayed an erosive step between test and control areas of enamel. and the pH of each solution increased slightly with increasing concentrations of CPP-ACP (pH 3. Abstract 2810 .80µm (SD 0. P with 0.125% CPP-ACP.25% CPP-ACP did not differ significantly (p>0.70). 0.Abstract 2810 Objectives: There have been numerous studies that have attempted to reduce the erosive potential of acidic beverages.06).063% CPP-ACP:1. 0.61.90). DDW: 0.063% CPP-ACP.31). P alone and P+0. 1.125% and 0.19µm (SD 0. All enamel specimens immersed in P with or without CPP-ACP exhibited red superficial staining.125%. 32 .25% CPP-ACP for 30 minutes at 37°. Enamel surface characteristics were examined qualitatively (stereomicroscopy. 3. 1.05).GC MI Paste – Abstract booklet – October 2007 57.07. 1.36). Previous studies have demonstrated the erosivity of Powerade™ and the purpose of this study was to determine the lowest concentration of casein phosphopeptide .34µm (SD 0. P+0.063%.40.07).63. Conclusion: The erosive potential of Powerade™ was eliminated or reduced on addition of low concentrations of CPP-ACP (0.09% and 0. 1. The titratable acidity decreased on addition of CPP-ACP (P=1. Messer LB andReynolds EC.87µm (SD 0.06).09%.09%. P+0.56). scanning electron microscopy) and quantitatively (surface profilometry).25% CPP-ACP: 0. P+0. IADR. 0. The mean profile depths were: P: 3. The mean erosive depths of DDW. San Diego 2002 . 0.

completed the 12-month examination. The baseline characteristics of the 2 groups did not differ. respectively). Participants included individuals who had had radiotherapy for head and neck cancer (n = 82) and others with Sjögren's syndrome (n = 56). The participants with the highest incidence and increment were those with Sjögren's syndrome in the CD-CP group. Baseline data collection was followed by reexamination 12 months later. Results: A total of 124 participants. a 33 . Proportionately more of the CD-CP group lost 1 or more teeth. but the difference was not statistically significant. 61 (49.8%) in the CD-CP group. although confirmation of this study's findings in other settings is warranted before a definitive conclusion can be reached. the small difference in coronal caries increment between the 2 groups was not statistically significant (0. Conclusion: It appears that CD-CP preparations hold promise as caries preventive agents for individuals with dry mouth. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2002.4 and 0. Posterior bite-wing radiographs were taken on both occasions.05% sodium fluoride mouthrinse among individuals with dry mouth.4% and 27%. Coronal caries incidence was higher in the sodium fluoride group than in the CD-CP group (34.3 surfaces. respectively).93:271-5 Objective: The purpose of this study was to compare the caries preventive efficacy of a mouthrinse solution containing casein derivatives Study coupled with calcium phosphate (CD-CP) with that of a 0. Some of that difference was accounted for by differences in baseline caries status. and the mean number of tooth loss was higher. Similarly. A clinical trial of the anticaries efficacy of casein derivatives complexed with calcium phosphate in patients with salivary gland dysfunction Hay KD and W. Design: A randomized control trial design was used.2%) in the sodium fluoride group and 63 (50.GC MI Paste – Abstract booklet – October 2007 58. Murray Thomson. There was insufficient root surface caries experience between the 2 groups observed for differences to be examined.

phosphate.8. GC Corp). mutans. the specimens were immersed in ion exchanged water (IEW) or were stayed in air of 100% RH at 37 degree Celsius.Caries prevention potential of a tooth-coating material containing casein phosphopeptideamorphous calcium phosphate (CPP-ACP) Sato T.GC MI Paste – Abstract booklet – October 2007 59.75mM. % diluted solution of TM and IEW was added to suspension of Streptococcus mutans (pH=4. Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) nanocomplexes have been shown to prevent demineralization and promote remineralization of enamel subsurface lesions in animal and in situ caries models. Conclusions: The results show that TM is effective in preventing the enamel demineralization in vitro and has strong butter capacity to acid produced by S. Göteborg 2003 . J Dent Res 2003 Nov.Abstract 1007 Objectives: The purpose of this study was to evaluate the caries prevention potential of a newly developed tooth coating material in vitro. Except those treatments. Incorporation of 1. The release of CPP-ACP and fluoride from the CPP-ACP-containing GIC was associated with enhanced protection of the adjacent dentin during acid challenge in vitro. Dashper SG. The aim of this study was to determine the effect of incorporating CPP-ACP into a self-cured glass-ionomer cement (GIC). P: 0. The extent of demineralization of the enamel specimens was determined by Knoop hardness measurements. 51. They were immersed in 10 wt. Ca: 0. 60. Yamanaka K and Yoshii E. Incorporation of casein phosphopeptide-amorphous calcium phosphate into a glass-ionomer cement Mazzaoui SA. ∆KHN of TM. The hardness reduction (∆KHN) was chosen as the primary efficacy viable. Tyas MJ.3. % diluted solution of each material for 10 minutes. IADR.75. 24. TM showed less Knoop hardness reduction (p<0. the pH of TM and IEW showed 6.2 and 30.82(11):914-8. FP and PP were 22.6 and 59.45mM) twice a day for 4 days. Abstract 1007. Two hours after the addition of samples. Methods: A newly developed tooth coating material: Tooth Mousse (TM. Buffer capacity of TM was also measured by pH monitoring. followed by 10 minutes immersion in demineralization solution (pH=4.0 respectively. MALDI mass spectrometry also showed casein phosphopeptides from the CPP-ACP nanocomplexes to be released. Bovine enamel specimens were prepared and assigned to 6 test treatments using 10 specimens per treatment group. Results: In IEW immersing group.56% w/w CPP-ACP into the GIC significantly increased microtensile bond strength (33%) and compressive strength (23%) and significantly enhanced the release of calcium.5) and the pH of suspension was monitored for 2 hours. Reynolds EC. a CPP-ACP free placebo (PP) and fluoride (900ppm F) added placebo (FP) were prepared. Burrow MF. ∆KHN of TM. 34 . General Session. Compared with FP and PP. and fluoride ions at neutral and acidic pH. Eakins D.05).0 and 4.6 respectively and in air staying group.6 respectively. Ten vol. FP and PP were 9.

relative to the control sugarfree lozenge. Conclusion: This study demonstrates that lozenges are a suitable vehicle for the delivery of CPP-ACP to promote enamel remineralization. human-enamel. Shen P. Walker GD.4mg (3 per cent w/w) CPP-ACP.8 and 56. without chewing. The CPP could be detected in plaque extracts 3 hrs after subjects chewed the CPP-ACP-containing gum. Shen P. After each treatment period the enamel slabs were removed. and the CPP were immunolocalized to the surfaces of bacterial cells as well as the intercellular matrix. Reynolds EC.8mg (1 per cent w/w) CPP-ACP. J Dent Res 2003 Mar. sectioned and subjected to microradiography and computerassisted densitometric image analysis to determine the level of remineralization. In the mouthrinse study. The aim of this study was to determine the effect of CPP-ACP incorporation into a sugar-free lozenge (pressed mint tablet) on enamel remineralization in a human in situ model.4mg of CPP-ACP increasing remineralization by 78 and 176 per cent respectively. Morgan MV. to be retained in supragingival plaque and remineralize enamel subsurface lesions in situ when delivered in a mouthrinse or sugar-free gum in randomized. Methods: The study utilized a double-blind. randomized. half-slab insets containing subsurface lesions. with that of other forms of calcium. the gum containing the CPP-ACP. only the CPP-ACP-containing mouthrinse significantly increased plaque calcium and inorganic phosphate levels. animal and human in situ caries models. produced the highest level of enamel remineralization independent of gum-chewing frequency and duration. 35 . and (iv) a no lozenge nil-treatment control. Aust Dent J 2003 Dec.48(4):240-3 Background: The anticariogenic potential of casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP) has been demonstrated using laboratory. Ten subjects wore removable palatal appliances with four. paired with their respective demineralized control. cross-over design with four treatments: (i) a lozenge containing 56.82(3):206-11 Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) nanocomplexes incorporated into sugar-free chewing gum have been shown to remineralize enamel subsurface lesions in situ. The aim of this study was to compare the ability of CPP-ACP.GC MI Paste – Abstract booklet – October 2007 61. four times per day for 14 days duration. 62. Cai F. (iii) a lozenge not containing CPP-ACP. double-blind trials. (ii) a lozenge containing 18. although not containing the most calcium per piece of gum. Results: The incorporation of CPP-ACP into the lozenge significantly increased enamel subsurface lesion remineralization with 18. embedded. In the chewing gum studies. Remineralization of enamel subsurface lesions in situ by sugar-free lozenges containing casein phosphopeptide-amorphous calcium phosphate Cai F. Lozenges were consumed. Retention in plaque and remineralization of enamel lesions by various forms of calcium in a mouthrinse or sugar-free chewing gum Reynolds EC. The results showed that CPP-ACP were superior to other forms of calcium in remineralizing enamel subsurface lesions.

This anticariogenic effect has been attributed to the multiphosphoseryl-containing sequences of casein and their ability to stabilize calcium phosphate. A substantial volume of literature now exists demonstrating an anticariogenic effect of dairy products (milk. Accepted for publication Paed Dent (not published) 64. J. animal and human in situ caries models. 58. Aust. which stabilise amorphous calcium phosphate (ACP) in solution by the formation of CPP-ACP nanocomplexes. Dairy components in oral health Reynolds EC. CPP-ACP is now being used commercially as an ingredient (Recaldent ) in oral care products. 36 . The CPP-ACP localise at the tooth surface and prevent demineralisation and promote TM remineralisation of enamel subsurface lesions. 79-81 Dental caries (tooth decay) is the localized destruction of tooth tissue initiated by specific dental plaque bacteria that ferment dietary sugar to organic acids.GC MI Paste – Abstract booklet – October 2007 63. Messer LB. Even though in most developed countries the prevalence of dental caries has decreased through the use of fluorides. Dairy Technol. the disease remains a major public health problem. powders and cheeses) in laboratory. milk concentrates. Erosion of human dental enamel by sports drinks Ramalingam L. Reynolds EC. These sequences can be released enzymically as casein phosphopeptides (CPP).

The results show that sugar-free gum containing CPPACP is superior to an equivalent gum not containing CPP-ACP in remineralization of enamel subsurface lesions in situ with mineral that is more resistant to subsequent acid challenge.38:551-556 The aim of this clinical study was to investigate the acid resistance of enamel lesions remineralized in situ by a sugar-free chewing gum TM containing casein phophopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP: Recaldent ). 37 . randomized. 2003. After each treatment the enamel slabs were removed and half of each lesion challenged with acid in vitro for 8 or 16 h.8%. of deposited mineral. Shen P. 2002). Reynolds C.com). Reynolds EC. Farrell et al. The casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) technology has been commercialized by the food and oral care industries as agents for the prevention and repair of early stages of dental caries (tooth decay) (Reynolds. Walker G. Cross KJ. while for the CPP-ACP remineralized lesions the corresponding reductions were 30.and 16-hour acid challenge of the lesions remineralized with the control gum resulted in 65.5 and 41. indicating that the remineralized mineral was more resistant to subsequent acid challenge. The study utilized a doubleblind.4 and 88.GC MI Paste – Abstract booklet – October 2007 65. Cai F. 1999. The 8.8 mg of CPP-ACP. crossover design with two treatments: (i) sugar-free containing 18.recaldent. Molecular modeling of the multiphosphorylated casein phosphopeptide αS1-casein (59-79) based on NMR constraints Huq NL. Reynolds EC. 1988). Reynolds et al. Acid resistance of enamel subsurface lesions remineralized by a sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) Iijima Y.0% reductions. The gum containing CPP-ACP produced approximately twice the level of remineralization as the control sugar-free gum. and (ii) sugar-free gum not containing CPP-ACP as control. 2000) (http://www. Journal of Dairy Research (2004) 71 28-32 Introduction: Casein micelles contain stabilized amorphous calcium phosphate that is bioavailable to the neonate (Holt & Sawyer. 66. respectively. 1996. Tryptic phosphopeptides formed from casein digestion associate with amorphous calcium phosphate forming stable nanocomplexes that have been described as calcium phosphate delivery vehicles (Holt et al. Caries Res 2004. The acid challenge after in situ remineralization for both control and CPP-ACP-treated lesions resulted in demineralization underneath the remineralized zone. The level of remineralization was determined using microradiography. Subjects wore removable palatal appliances with inserts of human enamel containing demineralized subsurface lesions and chewed the gum for 20 min 4 times per day for 14 days.

Huq NL. The αS1-CN(59-79) peptide stabilized calcium fluoride phosphate as amorphous nanocomplexes with a hydrodynamic radius of 2.5mM.9mM) was prepared for each material. Bovine enamel specimens were cut out and early caries lesions were formed in acetic acid buffer solution (pH4. In an approach to determine the ultrastructure of the casein phosphopeptide-amorphous calcium fluoride phosphate complexes. nine phosphate and three fluoride ions. nd 38 . Ca 1. have the potential to provide superior clinical efficacy in preventing dental caries and treating and repairing early stages of disease.Fluoride effect on acid resistance capacity of CPP-ACP containing material Kariya S.26nm. generation of fluoroapatite and accelerate remineralization.05). Recently. Sequence-specific resonance assignments were characterized by sequential (i. The spectral data were compared with that of the peptide αS1-CN(59-79 bound to calcium ions. Abstract 2045 . GC Corp.7 to 5.4 respectively. Methods: A CPP-ACP containing material (Tooth Mousse: TM. Reynolds EC. These casein phosphopeptides (CPP). form calcium phosphate delivery vehicles that retard enamel demineralization and pomote remineralization. the pH of TM and TMP were reduced from 7. 82 General Session. Thirty wt. Ten µL of 1N hydrochloric acid solution was continuously added to 10mL of 10wt. Specimens were immersed in each diluted material at 37 degree Celcius for 10 days. we have studied the structure of the predominant peptide αS1-CN(59-79) bound to ACFP using nuclear magnetic resonance (NMR) spectroscopy and X-ray diifraction. i +1).8.9 and 5. phosphate and fluoride delivery vehicle-αS1-casein (59-79) by stabilized amorphous calcium fluoride phosphate nanocomplexes Cross KJ.) and a second material containing 900ppm fluoride (TMP) were prepared. Yoshii E. Sum M.% diluted solution of TM and TMP. NMR studies of a novel calcium. Ca:1. After the addition of 100µL hydrochloric acid solution.5mM. P:0. i +2) nOes and Hα chemical shifts. The nanocomplexes exhibited stoichiometry of one peptide to 15 calcium. The neutralizing capacity of TM was a little superior to TMP. HEPES: 20mM.5) for 3 hours. mediumrange (i. 68. These complexes designated CPP-ACFP.12±0. Results: The amount of calcium loss of TMP and TM was 8. IADR. These effects would assist the improvement of acid resistance of TMP. we have shown that these peptides also stabilize calcium fluoride phosphate as soluble complexes.9) respectively and TMP showed lower levels of calcium loss compared woth TM (p<0. Honolulu 2004 . revealing that the structurally significant secondary NH and α-chemical shifts were similar.6) and 21.86µg/mm² (SD:1. Sato T. Stanton DP. % dilution by remineralizing solution (pH7.9mM) for 3 days. Biomaterials The repair of early tooth enamel lesions has been recently demonstrated by tryptic phosphopeptides derived from milk caseins that associate with amorphous calcium phosphate (ACP) forming stable complexes. containing the cluster sequence-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-. Neutralizing capacity of each sample was also measured by pH monitoring during acid addition. Fluoride has the capacity to improve the crystalline tooth structure. Sakaguchi Y. The acid resistance of the specimen was determined by measuring the amount of calcium loss after the immersion in lactic acid buffer solution (pH4. P:0.7µg/mm² (SD:1.Abstract 2045 Objectives: The purpose of this in vitro study was to evaluate the effect one acid resistance and neutralizing capacity when fluoride is added to a CPP-ACP containing material.GC MI Paste – Abstract booklet – October 2007 67.

. The group E (57±7µm) had significantly lower ld values compared to the group D (84±12µm).05). The remineralized enamel slabs without the wax were dehydrated and embedded in polyester resin.. G and H showed significantly lower ld values by 35-46% compared to the group A (105±16µm.3. of cheese. H: milk containing 0.3. ns). SD=703. while we failed to find the same ability in CO (X=393. Morita M. Honolulu 2004 . F: milk (Meiji Dairies Co. D: 30% soln. SD=705. 82 General Session. 0.2) was significant (p <0. Japan) G: milk containing 0. Methods: The enamel blocks prepared from bovine incisors were demineralized by immersion in a 0. Results: Paired t-test demonstrated that the remineralization ability for each GF (X=1682. µm). Subsequently.5 mM Ca.1 M lactic gel containing 6wt% carboxymethylcellulose at 37°C for 2 w. of processed cheese (Maiji Dairies Co. C: 10% soln.0) for 2 days at 50. which was standardized with NIH image. Objectives: To investigate the effect of xylitol chewing gums on remineralization of enamel in vitro.aC. xylitol chewing gum containing casein phosphopeptide-amorphous calcium phosphate (CP). IADR.01 M acetate buffer (pH4. E: 30% soln. Tukey’s test showed that the remineralization power for GF was significantly higher than that of CP and CO (p<0.4) and CP (X=780.0 mM CaCl2’ 0. were partially covered with wax for preserving demineralized surface. Conclusions: In conclusion.GC MI Paste – Abstract booklet – October 2007 69. In this study.05). 82 General Session. Tanaka T. For remineralization process. These treatments were repeated for 7 d and. p<0.0.5% CPP-ACP. of cheese containing 0. Paired t-test was conducted to analyze the remineralization ability of each xylitol chewing gum. Oda M. C. and experimental chewing gum only containing xylitol (CO) were used. finally.1 m NaCl:pH 7.5% CPP-ACP. E.5% CPP-ACP. Ikeda T. effects of processed cheese and milk containing CPP-ACP on enamel remineralization were examined in vitro. Miyake R. pH 7) for 23hr. nd 70. Koganei M.9 mM phosphate.2. IADR.3 with KOH) containing extracts from each chewing gum. Honolulu 2004 .Abstract 2049 Objectives: It has been known that a sort of food such as cheese can enhance remineralization. Abstract 2046 – Remineralization power by xylitol chewing gums Murata Y. Each 6 demineralized enamel samples were exposed to one of the following solutions for 60 min: A: untreated (baseline). B: 10% soln. it was suggested that addition of CPP-ACP into processed cheese or milk would be effective for enhancement of remineralization in enamel lesions.2% CPPACP. Komatsu H. Methods: Xylitol chewing gum containing Gloiopeltis furcata extract and calcium hydrogenphosphate (GF).Abstract 2046 It is considered that chewing gums containing some additives promote remineralization of enamel. the samples were immersed in the mineral solution (1. which were prepared by immersing in 0.aC in a remineralizing solution (1. Results: The groups B. Remineralization power was analyzed quantitatively by contacting microradiography for mineral change (vol%xµm) between demineralized and remineralized surface. Inaba D. Approximately 100µm-thick enamel sections were prepared. Matsuda Y. Sano H. of cheese containing0. enamel slabs with the wax were immersed for 14 days at 37 .. Japan). Yonemitsu M. So did the group G (64±10µm) and H (63±12µm) with respect to the group F (91±6µm) indicating improved mineral recovery.6mM KH2PO4’ 0. One-way ANOVA and Tukey’s test were then conducted between each independent variable. Twenty-four demineralized enamel slabs. Conclusion: The remineralization power of chewing gums differs depending on the additives the chewing gum contained. the samples were microradiographed to measure the lesion depth (ld..05). nd 39 . SD=667. Abstract 2049 – Effects of cheese and milk containing CPP-ACP on enamel remineralization Minami K. 20 mM Hepes.

On the other hand. These results correlate well with the ability of the CPP to bind to both amorphous and crystalline phases of calcium phosphate. Therefore we focused on the ultrasonic pulse method and tried to identify the condition of demineralization with time. Huq NL. nd 72. using lowspeed hard tissue precision cutting machine (Isomet 1000. Hinoura K. 2) Storage in the demineralizing solution The specimens were stored in the 0.1M lactic acid buffer solution for 10 and 30 minutes. Determination of demineralization of tooth substrate by use of an ultrasonic device. it was suggested that the condition of demineralization of the tooth structure could possibly be measured non-destructively by using ultrasonic pulse method. Ando S.Abstract 2997 Objective: The repair of early enamel lesions has been demonstrated by tryptic phosphopeptides derived from milk caseins that associate with amorphous calcium phosphate (ACP) forming stable complexes. dynamics and simulated annealing. therefore. which occurred in longitudinal wave. Panametrics) as an ultrasonic transmitter/receiver. Each measurement was conducted at 23 ± 1 . the calcium phosphate delivery vehicle aS2CN(2-20). and every 7 days until 28 days after starting the storage in the solution.0). together with the changes in the conformation along the backbone as revealed by the Ramachandran plots and orientation of the side-chains indicate the presence of turns and loops with a high degree of flexibility between these secondary structural elements. it could be considered that the inorganic component of the tooth structure dissolved by the demineralizing solution. 50 ± 5%RH in the temperature-controlled room. Conclusion: From the result of this study. Takamizawa T. and stored in the artificial saliva (pH 7. All calculations were peformed with the molecular package SYBYL 6. Other specimens were stored in the 10-times diluted solution of CPP-ACP Paste (Tooth Mousse. All the surfaces of the block were polished accordingly to the degree of a water-resistant SiC paper #2. Above measurement was conducted before the storage in the solution every day for one week. This block was used as the measuring specimen. Method: 1) Fabrication of the specimens for the measurement The extracted bovine lower anterior teeth without crack or deficiently mineralization. the thickness of each specimen and its longitudinal wave sonic speed was measured. The number of specimens used was 6 pieces for each condition. The specimens were prepared by cutting the enamel and dentin of labial surface of the above bovine teeth in a block (4x4x1mm). they were also stored in the 0. Reynolds EC. no decrease in the sonic speed was found for both of enamel and dentin specimens stored in the 10-times diluted solution of CPP-ACP paste and even an increase of the sonic speed was found especially for the enamel. 3) Measuring of the propagation time of ultrasonic waves Pulser/Receiver (MODEL 5900. 40 . Method: The aim of this project was to characterize. which means. in order to establish an in-vitro model of demineralization or remineralization. The specimens were stored in the demineralizing solution twice a day. Buehler). was measured by ultrasonic transmission method. The in silico peptide aS2-CN(2-20) was constrained using distance constraints obtained from nuclear magnetic resonance (NMR) spectroscopy data. the changes of the sonic speed. form calcium phosphate delivery vehicles that retard enamel demineralization and promote remineralization. After that. Miyazaki M. Oscilloscope (Wave Runner LT584. The specimen was settled on the stage and contacted by the transducer. Honolulu 2004 . 82 General Session. it is considered as important to determine the condition of demineralization on the tooth structure in order to prevent a caries formation to be caused as a result of the demineralization. using molecular modeling techniques. This tendency to decrease of the sonic speed meant the decrease in the inorganic component of the tooth structure.1M lactic acid buffer solution and then in the artificial saliva at 37 . These casein phophopeptides (CPP). Inage H. Next. by measuring the changes on the tooth structure. IADR. at the age form 2 to 3 years old were used for this trial. Results: The RMSD values. Accordingly. Japan J Conserv Dent 2004 Jun 47 Spring Issue 24 – Abstract B-4 Objective: It has been proven that a tooth structure is maintaining its dynamic balance by repeating demineralization and remineralization. It could be considered that the inorganic component contained in high concentrations in CPP-ACP acted to prevent demineralization or to enhance remineralization of the tooth structure. Each propagation time of ultrasonic waves. Results: The ultra-sonic speed of enamel and dentin specimens was found to decrease with time for a specimen stored in the demineralizing solution compared to a control specimen stored in the artificial saliva. Cross KJ.000. Yamaguchi K.GC MI Paste – Abstract booklet – October 2007 71. fifty conformers were produced that were stereochemically correct with low nOe violations indicating good consistency with the nmr data.8 on a Silicon Graphics Octane workstation. Lecroy) and a specimen stage were used as the measurement system. After energy minimization. Conclusion: In conclusion molecular modeling is an essential tool in the investigation of the anticariogenic mechanism of the CPP because it enables the visualization of the peptide conformational preferences and allows the identification of structural features that are important in its role as a calcium phosphate delivery vehicle. Abstract 2997 – Molecular of anticariogenic casein phosphopeptide aS2-CN(2-20) NMR spectroscopy derived constraints Deangelis EF. GC) and placebo paste (no CPP-ACP contained) for 10 minutes prior to the storage of the specimens in the demineralizing solution.

Conclusion: Out of this clinical study. Braem M. GC Tooth Mousse appears to be effective in reducing tooth hypersensitivity.9 (U). Approval for the study was obtained from The University of Melbourne Human Research Ethics Committee. sensitivity of the test teeth was scored by the dentist on a 0-10 visual analogue scale (VAS) after air-blast (A). animal and human in situ experiments. 8% remineralization by the 220 ppm F rinse. Reynolds C. Reynolds EC. 41 . Australian Dental Journal ADRF Special Research Supplement 2004. oral hygiene and dietary habits were recorded before treatment. A significant reduction (Wilcoxon signed rank statistics. Each treatment was for 14 days duration and at the completion of each treatment the enamel half-slabs were removed.4% CPP-ACP and 220 ppm F produced 19% enamel subsurface lesion remineralization compared with 2% remineralization produced by the placebo rinse. Poitevin A. (iii) 220ppm F . All subjects were instructed to insert the intra-oral appliance immediately before mouthrinsing for 60 seconds and to wear the appliance for a further 20 minutes. Walker G.4% CPP-ACP. Coutinho E.30 pm. 74. Cai F. leave for 3 min. Clinical effectiveness of a CPP-ACP crème for tooth hypersensitivity treatment. 19-67 yr age range) for tooth hypersensitivity using GC Tooth Mousse. Medical and dental history. After 3 weeks.6±2. The results support the role of fluoride in promoting remineralization and demonstrate an important facilitation of the effect of fluoride by CPP-ACP. Van Landuyt K.3±2.1±3. 25-28 August 2004.0±2. paired with their respective demineralized control. Van Meerbeek B. stains and fluorosis were selected from extracted human third molars and polished wet to a mirror finish using SoftexTM (3M) discs. Objectives: To evaluate the clinical effectiveness of a newly developed tooth-coating crème (GC Tooth Mousse). De Munck J. and 14% remineralization by the 0. The study utilized a double blind. P. 11.7 (A).GC MI Paste – Abstract booklet – October 2007 73.8.2±2. p<0. Each polished surface was then sawn from the tooth as an approximately 8 x 4 mm slab and lesions were created in the enamel windows using a lactate pH 4. Abstract 0136 Tooth hypersensitivity is a widespread oral discomfort that remains difficult to treat. 24 male.8 (A). 3.3 (U). (ii) 0.30 am. Nine healthy subjects wore removable palatal appliances with four half-slab insets of human enamel containing demineralized subsurface lesions. containing Casein PhosphoPeptide .Amorphous Calcium Phosphate (CPP-ACP) for the treatment of tooth hypersensitivity. All these values were significantly different to each other (p<0. This was performed four times per day at the following times: at 10 am.49:4 Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) have been demonstrated to have anticariogenic activity in laboratory. Sensitivity of the test teeth was daily evaluated by the patient using VAS. Methods: 11 private practitioners treated 61 patients (37 female.3±2. Regarding the daily evaluation of tooth sensitivity by the patient. sectioned and subjected to microradiography and computer-assisted densitometric image analysis to determine the level of mineral content. patients sensitivity was re-scored by the dentist (A. The mouthrinse containing 0. and U). The change in sensitivity was more apparent after the air stimulus than the tactile stimuli. apply GC Tooth Mousse in tray or with swab.0001) in tooth sensitivity was noticed by the dentist after 21 days to 3. EADR Istanbul. Enamel remineralization by a mouthrinse containing casein phosphopeptide-amorphous calcium phosphate and fluoride in an in situ model. randomized. 2. and (iv) mouthrinse containing neither CPP-ACP nor fluoride (placebo). Peumans M. the sensitivity also decreased from day 1 (5.4) to day 21 (2.9±2. Half of the patients reported a general reduction in sensitivity (44%) and wanted to repeat the treatment if sensitivity re-occurred (43%).0±2. four-way crossover design with four treatments: (i) 0. instrumentation with a probe (P) and an ultrasonic scaler (U).4% CPP-ACP and 220 ppm F . The other half of each enamel slab was stored in a humidified container and was used as the control demineralized lesion. 2 am and 3. At baseline. Sound relatively planar buccal and lingual surfaces free of cracks.01). spread remaining crème throughout the mouth for 1-2 min.3). embedded. Shen P.9 (P) and 5. carbopol demineralization buffer. the test teeth showed a VAS sensitivity of 6. Results: Before treatment. The patient was instructed to apply GC Tooth Mousse during 21 days (brush teeth at evening.4% CPPACP rinse. The aim of the current study was to compare the enamel remineralization ability of mouthrinse containing CPP-ACP with that of a mouthrinse containing fluoride in an intra-oral enamel remineralization model.4 (P) and 4. do not eat or drink for 30 min).

To date the potential anti-cariogenicity of the CPP-ACP complex has been demonstrated in a rat caries model. p111-118 For details ref. 2001. Investigation of the binding of casein phosphopeptides to the major enamel pellicle proteins. Parotid saliva was collected using a Lashley cup after stimulation with lemon drops. in situ human caries models. Flat bottomed. However. depressing enamel demineralization and enhancing enamel remineralization of early lesions. Textbook: Preservation and Restoration of Teeth . Reynolds EC. The anticariogenic mechanism of the casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) complexes has not been fully described. The profiles of intact untreated saliva were examined by SDS-PAGE. The plate was washed and 50µL of polyclonal rabbit antibovine casein antibodies (1:640) in 0. All peptides contain the sequence motif –Ser(P)-Ser(P)-Ser(P)-Glu-Glu-. Cross KJ. The major casein phosphopeptides (CPP) are αs1 -CN(59-79) and β-CN(1-25) and its deamidated forms with smaller amounts of αs2 –CN(1-21) and αs2 –CN(46-70). Additional aids to the remineralisation of tooth structure. where Ser (P) denotes an Ophosphoseryl residue. Australian Dental Journal ADRF Special Research Supplement 2004. Oral cavity surfaces are constantly exposed to free flowing saliva. 2003). 1991. Similarly the profiles of the salivary proteins bound to HA were also examined by SDS-PAGE and MALDI-TOF mass spectrometry for comparison with previously published reports. The ELISA revealed that both αs1 –CN (59-79) and β-CN (1-25) bind to whole and parotid saliva. Cai et al. Walsh LJ.5% BSA and incubation for three hours at RT. Textbook: Preservation and Restoration of Teeth . HPLC and MALDI-TOF mass spectrometry.. The purity was checked by MALDI-TOF mass spectrometry.5% (w/v) BSA was added and incubated overnight at RT.GC MI Paste – Abstract booklet – October 2007 75. The plate was washed six times and 100µL of substrate solution was added to each well. Six washes were performed followed by the addition of 100µL of horse radish peroxidase-conjugated goat anti-rabbit antibody (1:2000) in 0. Ung M. Previous studies have shown that CPP-ACP is incorporated into dental plaque. 76. Superimposition of the binding curves suggested that β (1-25) casein had a greater degree of binding to whole saliva.. Reynolds EC. Upon contact with saliva. Therefore. It has been proposed that CPP localize ACP in dental plaque. the interactions between CPP and salivary proteins has not been previously investigated.. Huq NL. The plate was washed four times with PBST. The aim of this study was to investigate the binding of the CPP to salivary proteins using an enzyme-linked immunosorbent assay (ELISA). binding of the anti-bovine casein antibodies to whole and parotid saliva was tested and confirmed to be negligible. using the three letter codes for the amino acryl residues.2nd edition 2005. The sequences are shown below. This interaction may represent a crucial phase in the anticariogenic activity of CPP. After 15 minutes of development the optical density was measured at 415nm. Since the salivary statherin is also multi-phosphorylated and was postulated to be delivered from the ancestral milk casein. p111-118 42 . 96-well polyvinyl microtitre plates were coated with 50µL of clarified whole saliva (10µg/mL) in PBS and incubated at 4°C overnight. Shen et al. tooth enamel is immediately covered by a thin layer of salivary proteins that forms the acquired enamel pellicle. in vitro remineralization/demineralization models and in shortterm mouthwash trials (Reynolds. thus acting as a reservoir of free calcium and phosphate. understanding the exact mechanism of CPP activity may enable its use in a greater number of therapeutic applications and lead to further improvement of its specific activity and efficacy. followed by the serial dilution of CPP (5mg/mL) in PBS and incubation for three hours at RT. et al.49:4 Tryptic phosphopeptides derived from milk caseins are known to associate with amorphous calcium phosphate (ACP) forming stable complexes that behave as calcium phosphate delivery vehicles and are effective in the remineralization of early enamel lesions. An ELISA was developed using polyclonal rabbit anti-bovine casein antibodies to detect the CPP. and it has been suggested that the hydrophobic residues in CPPs may be involved as attachment anchors at hydrophobic sites in plaque and oral tissues. Reynolds. 1995. this is the first time the direct binding of the anticariogenic CPP to salivary proteins has been demonstrated. Whole unstimulated saliva was collected from adults.2nd edition 2005. However. The coating solution was removed and the wells blocked with 200µL of 1% (w/v) BSA in PBST for one hour at RT. + The casein phosphopeptides αs1 –CN (59-79) and β-CN (1-25) were selectively precipitated from a tryptic digest of casein using Ca² and ethanol and further purified by anion exchange FPLC and reversed phase HPLC.

at the clinical level. The gum containing CPP-ACP produced approximately twice the level of remineralization as the control sugar-free gum. of deposited mineral. a clinical case is presented which allows us to stress the importance of the complementary use of prophylactic and restorative measures. Reynolds EC. Walker G. and the patients who have the greatest risk of developing caries may not always benefit from optimal fluoridation. The capacity of the glass ionomer cement restorations to remineralize the dental tissues internally and externally is also discussed. The treatment with ozone is debated and illustrated in a case of remineralization of cervical lesions in a patient under neuroleptic medication.8 mg of CPPACP. 2004 pp.GC MI Paste – Abstract booklet – October 2007 77. and joint therapies with these are proposed. if carious lesions must be intercepted at an early stage.8%.and 16-hour acid challenge of the lesions remineralized with the control gum resulted in 65. in principle. respectively. and (ii) sugar-free gum not containing CPP-ACP as control. Martin JM. indicating that the remineralized mineral was more resistant to subsequent acid challenge. Subjects wore removable palatal appliances with insets of human enamel containing demineralized subsurface lesions and chewed the gum for 20 min 4 times per day for 14 days.261-275 Despite the widespread diffusion of topical fluorides.15 n°3.0% reductions. 78. These findings justify focusing on therapeutic agents which are capable of reacting jointly with fluorides. none of them can substitute for optimal topical fluoride application. The 8. to develop dual strategies of remineralization and restoration. It is shown that even if the prophylactic measures using these agents and materials show undeniable clinical importance. 20% of those patients who do not. Miller C. belong to a risk group still present with carious lesions. The roles of chlorhexadine. La reminéralisation des lesions carieuses (2) synergies thérapeutiques / The remineralisation of caries lesions: joint therapies Lasfargues JJ. crossover design with two treatments: (i) sugar-free gum containing 18. Caries Res 2004. Cai F. The level of remineralization was determined using microradiography. Reynolds C.4 and 88. xylitol. Realites Cliniques Vol. randomized. In addition. while for the CPP-ACP-remineralized lesions the corresponding reductions were 30. In this article. After each treatment the enamel slabs were removed and half of each lesion challenged with acid in vitro for 8 or 16 h. and it is proper. Shen P. and caseine phosphopeptides on remineralization are discussed. in the patient at high risk the caries are already present. The results show that sugar-free gum containing CPP-ACP is superior to an equivalent gum not containing CPP-ACP in remineralization of enamel subsurface lesions in situ with mineral that is more resistant to subsequent acid challenge. By way of conclusion. The acid challenge after in situ remineralization for both control and CPP-ACP-treated lesions resulted in demineralization underneath the remineralized zone. Acid Resistance of Enamel Subsurface Lesions Remineralized by a Sugar-Free Chewing Gum Containing Casein Phosphopeptide-Amorphous Calcium Phosphate Iijima Y. 43 . the various therapeutic possibilities for remineralization that may be associated with fluorides are discussed.38:551-556 The aim of this clinical study was to investigate the acid resistance of enamel lesions remineralized in situ by a sugar-free chewing gum containing casein phosphopeptideamorphous calcium phosphate nanocomplexes (CPP-ACP: Recaldent™). The study utilized a doubleblind.5 and 41.

) on demineralization by measuring changes in the ultrasound transmission velocity. It is also possible that CPP-ACP acted to prevent demineralization or to enhance remineralization of the tooth tissues. followed by 10 min immersion in a demineralization solution (pH 4. no decrease was found for specimens stored in the CPP-ACP solution and a significant increase was found for enamel specimens. and then in artificial saliva (pH 7. Yamaguchi K. However. The buccal surface of each specimen was moistened for 5 min or 24h with 10µl of the control or one of the four test solutions Olaflur. Abstract 94 – 52 ORCA Congress. Indianapolis.1 M lactic acid buffer for 10 min. Martin JM. Realites Cliniques Vol.GC MI Paste – Abstract booklet – October 2007 79. July 2005. Other specimens were stored in a 10 x diluted solution of CPP-ACP paste and a placebo paste without CPP-ACP for 10 min. Takamizawa T. The specimens were blocks of bovine enamel and dentin. significantly more fluoride was associated with the superficial layer up to 20 µm. On the other hand. human third molars. Cubical enamel and dentin specimens were cut out from extracted. 80. Jung H.75. Hinou K. Miller C. The values were 3-4 times higher in enamel and 4-8 times higher in dentin after 5-min application time and 10-24 times higher than the initial fluoride content in both hard tooth tissues after 24-h application time. 44 . Focusing on the experimental solution. Oleaflur.75 mM. Hiller KA. Schmalz G. 2004 pp.05) were done. From the results of this study. The specimens were ground in 20µm steps and the fluoride content was determined in each enamel and dentin layer. Six specimens were used for each condition. The specimens were stored in 0. The ultrasound transmission velocity in specimens was found to decrease with time for specimens stored in the demineralization solution. Supported by a grant to promote multi-disciplinary research project. Using ultrasound transmission velocity to analyze demineralization of tooth substrate Lasfargues JJ. cariesfree.45 mM) twice a day before storage in artificial saliva. and to determine the effect of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste (Tooth Mousse.0). the fluoride levels in enamel and dentin were somewhere in the order of the values of sodium and amine fluoride solutions. or experimental fluoride containing hydrolyzed casein. sodium fluoride. it is suggested that the condition of demineralization of the tooth structure could possibly be measured non-destructively by using ultrasonic pulse method. After application of the fluoride solutions. USA / Caries Res 2005. Ca 0.and long-term application of four fluoride solutions.261-275 nd Miyazaki M. including a casein-based fluoride preparation. Inage H. and one-way ANOVAs followed by the Tukey HSD test (α = 0. Friedl KH. GC Corp. Fluoride uptake and distribution in enamel and dentin after application of different fluoride solutions Holler BE. P 0. a tendency towards higher values could be observed after application of the experimental solution.15 n°3. 39:319 The purpose of this study was to investigate the use of an ultrasonic pulse method to analyze demineralization of dental hard tissues. The propagation time of longitudinal ultrasonic waves was measured by a pulser-receiver with a transducer. Clin Oral Invest (2002) 6 :137-144 The aim of this in vitro study was to examine the fluoride accumulation in enamel and dentin after short.

Artificial root surface caries were created over a 10-day period (alternating 16 hours demineralization followed by 8 hours remineralization). and this may be an important factor in reducing susceptibility of root surfaces to a cariogenic challenge. Mean lesion depth reduction for CPP-ACP was 33% compared with NaF Rinse (P<. 2) NaF Rinse Group:216+/-21um.DMR). the placebo paste. Chen Li.05% NaF). MIP showed a significant protective affect against acid induced demineralization (p < 0. Results: Mean root surface lesion depths were: 1) No Treatment Control Group: 310 +/. fluoride paste and water controls were -1.05). when compared with fluoride rinsing (0.. ACT McNeil-PPC. ANOVA. 3) CPPACP Group: 144+/-19um. Sato T. Kariya S. Toshiba) analysis. A placebo paste without CPP-ACP. treated with 0. pH 5. Flaitz C. X-Ray CT further confirmed this result.50 respectively.27µm.05% sodium fluoride rinse (NaF. Methods: Bovine enamel specimens were prepared. Mean lesion depth reductions compared with the No Treatment Control Group were: 30% for NaF Rinse (P<.3 grams of MIP and then challenged with 10 ml of a demineralization solution (0. ANOVA. and -16. Results: QLF analysis determined that the mean change in fluorescence radiation (Δ F. n=12).Abstract 3275 Objective: This in vitro study evaluated the effect of a casein phosphopeptide-amorphous calcium phosphate paste (CPP-ACP) on artificial caries formation in human root surfaces using polarized light microscopy. -9. Conclusion: This QLF study confirmed the inhibitory effect of CPP-ACP on acid induced surface demineralization and demonstrated a functional role for MIP ™ as a caries preventative agent for use in minimum intervention dentistry. Inspektor Dental Care) before and after the procedure. The collected images were then analyzed to determine the degree of demineralization. which contains 10% Casein Phosphopeptide – Amorphous Calcium Phosphate (CPP-ACP). DMR) and 54% for CPP-ACP (P<. No treatment control root segments were exposed to synthetic saliva only for a 14-day period. Preventing acid induced enamel demineralization using CPP-ACP containing paste Sakaguchi Y.05. by use of Quantitative Light-Induced Fluorescence (QLF). The sections of enamel samples were also subjected to X-ray CT(TOSCANER-30000µhd. Compared to the placebo and fluoride paste. Bioavailable calcium and phosphate in CPP-ACP has been demonstrated to bind to hydroxyapatite. rd 82. ANOVA. GC America. Conclusions: Casein phosphopeptide-amorphous calcium phosphate paste markedly enhanced the resistance of root surfaces to artificial caries formation. DMR). and then exposed to synthetic saliva for 24 hours.55.05. rd 45 . using QLF (Inspektor pro. Each portion a single tooth was assigned to one of the three treatment groups: 1) CPP-ACP (MI Paste. and 3) No Treatment Control (n=12). IADR. -6.03. 83 General Session.Abstract 2055 Objectives: The objective of this in vitro study is to evaluate. CPP-ACP and NaF root segments were treated with the appropriate assigned agent for 60 seconds. Methods: 12 human teeth with sound root surfaces were sectioned from into 3 portions.98. a CPP-ACP free paste containing 900 ppm Fluoride and water were included as controls.05.GC MI Paste – Abstract booklet – October 2007 81. 83 General Session. IADR. 2) 0. n=12). CPP-ACP and NaF treatment. The enamel surface of each specimen was scanned.0) at 37 degree Celcius RH 100% for 72 hours. Casein phosphopeptide-amorphous calcium phosphate paste: root surface caries formation Hicks J. Baltimore 2005 . ANOVA. rinsing and synthetic saliva exposure were repeated on a daily basis over a 14-day period. the caries preventive potential of MI Paste (MIP. %) of the MIP. followed by air-water rinsing for 60 seconds. Kato S. DMR). GC Corp. also known as GC Tooth Mousse ™.1M Lactic acid buffer. Baltimore 2005 . Longitudinal sections (3 sections/root segment) were obtained and lesion depths were determined using polarized light microscopy (n=36 lesions/group.

Troianiello S. 46 . dental materials and equipment. This first in a series of 2 articles describes and illustrates oral disease management in geriatric MID. uses the medical model.GC MI Paste – Abstract booklet – October 2007 83.N°5 The aging of the population combined with increased retention of natural teeth into old age means that clinicians now face a new caries challenge in older dentate patients. Minimal intervention dentistry: part 1. Nuove strategie nella prevenzione della carie dentaria: studio sperimentale sui caseino-fosfopeptidi Ferrazzano GF. external and internal remineralization. and surgical intervention only when required and only after disease has been controlled. Ingenito A. a modern evidence-based approach to caries management in dentate patients. integrated with an evaluation of the plaque-biofilm interface and the resultant dynamic oracl disease process. which involves the assessment and management of a diverse range of primary and modifying factors. Vol. Minimal intervention dentistry (MID). Prevenzione odontostomatologica 2005. An increase in the onset of dental caries is evident among patients who may not have had high levels of caries in the past and who may have undergone extensive restorative procedures during their lifetimes. Strategies for addressing the new caries challenge in older patients Chalmers JM. with a focus on early detection and prevention. JCDA. use of a range of restorations.72. The main components of a geriatric approach to MID are assessment of the risk of disease. 4:15-21 84. whereby disease is controlled by the “oral physician” and an affiliated dental team. Jule 2006.

CPP-ACP in sugar-free gums has been shown to significantly slow the progression and promote the regression of caries in a randomized. which acts as a calcium and phosphate reservoir when incorporated into dental plaque and on the tooth surface. double-blind studies wore removable mandibular appliances with a demineralized human enamel sample. and significant enhancement of calcium.7% of the hypomineralised fluorotic lesion. Recently.Abstract 184 Objectives: This study examined the effect of a sugar free chewing gum (Tablet type) containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP. Results: During the second acid challenge. 8 Congress of European Academy of Paediatric Dentistry. significantly increasing after enamel pre-conditioning with NaOCl. phosphate and fluoride ions at neutral and acid pH. Amsterda. Advantages of increased remineralization are indicated for CPP-ACFP over CPP-ACP due to the availability of the fluoride ion with calcium and phosphate ions at the enamel surface. A chemical approach Manton DJ. lozenge. June 2006 – Abstract I4 Casein phosphopeptide – amorphous calcium phosphate (CPP-ACP. controlled clinical trial. Recaldent. The appliances were inserted in oral environment immediately before gum-chewing for 20 min and then retained for another 20 min. This was performed four times per day for 14 days. with a concomitant improvement in aesthetic appearance. Iijima S. The addition of low concentrations (<1. The CPP-ACP nano-complexes release calcium and phosphate ions via a pH or concentration gradient mechanism to maintain a supersaturated environment with respect to hydroxyapatite. 84 General Session. however setting time was increased.stabilized amorphous calcium (fluoride) phosphate. it seems that this gum remineralizes subsurface enamel with mineral of higher crystallinity than saliva. with added calcium and phosphate. The reason why there was a less mineral loss during the second acid attack for 3 days in case of CPP-ACP gum. Nishimura M. Brisbane 2006 . Methods: Twenty subjects in randomized. mouthrinse.0%) of CPP-ACP to acidic sports drink has been shown to reduce erosive potential without significantly altering taste. Clinically. The formation of higher crystallinity makes the mineral less soluble in acid. Remineralization with CPP-ACFP of mild fluorotic lesions of enamel in vitro resulted in remineralization of 44. The enamel samples were acid challenged for 3 days in vitro. therefore reducing demineralization and enhancing remineralization. toothpaste and added to GIC restorative material TM th 86. The difference between the two groups was statistically significant and the probability was 0. The addition of CPP-ACP to Glass Ionomer Cement (GIC) restorative material resulted in the inhibition of demineralization of adjacent dentine. The inclusion of fluoride into CPP-ACP (as CPP-ACFP) forms a novel material with fluoride incorporated into the nanocomplex. CPP-ACP can de delivered to the tooth surface in several vehicles: chewing gum. cross-over. Recaldent ) is a casein derived peptide. with increase in microtensile bond and compressive strength. increasing fluorapatite formation. th 47 .GC MI Paste – Abstract booklet – October 2007 85. Conclusion: Remineralized enamel by CPP-ACP tablet gum was much higher acid resistant than placebo gum. topical crème. Promoting remineralization: using casein phosphopeptide .0 ± 9. IADR. EAPD. The ability of CPP-ACP to remineralise white spot lesions of enamel (WSL) both in vitro and in situ has been widely reported. CASRN: 691364-49-5) on an acid resistance of remineralized enamel without fluoride conditions in a human in situ model. Intra-oral experiment using another gum was performed after 1 week of washout time. Acid resistance of remineralized enamel by a sugar-free chewing gum Iijima Y. the mineral loss level of CPP-ACP tablet gum group showed less mean mineral loss (-2142) than that of placebo gum (-2804).036. Half of the enamel samples were covered with nail varnish after intra-oral exposure for remineralization. sectioned and subjected to microradiography to determine the level of mineral vol%.

3% enamel subsurface remineralization.05).5% CPP-ACP at pH 5. Brisbane 2006 . sectioned and subjected to microradiography and computer-assisted densitometric image analysis to determine the level of remineralization. Reynolds C. 11:30am. This study was supported by the Cooperative Research Centre for Oral Health Science. There was a one-week washout period after which the subjects crossed over to the other mouthrinse. The % mineral profiles of each WSL and adjacent sound enamel were compared and differences between sound and lesion values calculated (ΔZd. randomized.9% remineralization of the enamel subsurface lesions in the 10 day period whereas the pH 7.5% CPP-ACP rinse at pH 7.Abstract 185 Objectives: To investigate the potential of a commercially available dental crème containing casein phosphopeptide – amorphous calcium phosphate (CPP-ACP). embedded.5 in remineralization of enamel subsurface lesions in an human in situ model. Shen P. IADR. Mineral content of each remineralized half-slab invested in resin with its control half slab was determined by microradiography after sectioning and lapping to 85 ± 5µm. Reynolds EC. Proportional change in mineralization (%R) was calculated according to the formula: %R = ((ΔZd−ΔZr)/ΔZd) x 100 and data analysed (ANOVA. The University of Melbourne.3 ± 2. Conclusion: A 20% dilution of Tooth Mousse crème containing CPP-ACP produced 551% more WSL mineralisation than a placebo crème in an in situ model. Remineralisation of white spot lesions in situ by tooth mousse Manton DJ.Abstract 189 Objectives: To investigate the efficacy of mouthrinse containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) at pH 5. 4ml H20) of Tooth Mousse (TMtest) or placebo Tooth Mousse (TMPlacebo) was placed intra-orally (60s) after appliance insertion. Healthy volunteers (n=6) wore mid-palatal appliances containing four enamel test slabs with WSL. cross-over design with two treatments: (i) a mouthrinse containing 0. 2-way cross-over in situ study.GC MI Paste – Abstract booklet – October 2007 87. Specimens were divided into test and control half slabs. 2:00pm and 3. th 88.2 ± 1.22 ± 3.5% CPP-ACP at pH 7. Method: This study utilized a double blind. After each treatment period the enamel slabs were removed. drinking and oral hygiene procedures.0 rinse resulted in a 10. A slurry (1g crème.10%. 84 General Session.5 Shen P. ["GC Tooth Mousse" (10% w/v CPP-ACP) (GC Corp. The appliance was worn (40min) and the process repeated 4 times per day for 10d. Results: Use of the mouthrinse containing 0. ΔZr). Cai F.5.30pm) for 10 consecutive days.5 produced 14. IADR. Messer LB.0 and (ii) a mouthrinse containing 0. Cai F. Results: TMPlacebo resulted in a %R of 3. p<0.5% CPP-ACP at pH 5. paired with their respective demineralized control. School of Dental Science. Remineralization by a mouthrinse containing CPP-ACP at pH 5. Reynolds C.5% CPP-ACP mouthrinse at pH 5. Reynolds EC. Cochrane NJ.72 ± 2.5 produced significantly greater (38%) remineralization in situ than a 0. th 48 . Methods: Enamel specimens were sectioned from either buccal or lingual surfaces of extracted sound human third molars and WSL windows were created using the Carbopol method. Walker GD. The subjects were instructed to rinse with 5 ml of mouthrinse for 60 seconds 4 times a day (at 10:00am. Brisbane 2006 . whereas TMtest produced %R of 24. Cross-over occurred after a 7d washout. Tukey's post-hoc.31%. Subjects wore removable palatal appliances with four human-enamel half-slabs inset containing subsurface demineralized lesions. 84 General Session. The subjects did not wear the appliances during eating. Conclusion: A 0. Japan)] to remineralize sub-surface white spot lesions of enamel (WSL) in a double blind.0.

Kato S. Toshiba) and Scion Image for Windows before and after intraoral treatments. (ii) sugar-free gum containing 20 mg citric acid alone. Methods: This cross-over in situ model involved five healthy adult subjects and four randomized treatments: tooth creme containing 10% CPP-ACP. 12. The aim of this study was to investigate the effects of CPP-ACP in a fruit-flavoured sugar-free chewing gum containing citric acid on enamel remineralization. th 90. TM. wear the appliance for 30 min. TMP was superior to all other formulations (p < 0. (Tooth Mousse.2%) or the gum containing citric acid alone (2. Reynolds C. Chen L.73 ± 11. pH 5.57 respectively. Brisbane 2006 . rinse the lesions with water. Manton DJ. Results: The mean percentage remineralization for the placebo. Walker GD.14 ± 14.8 mM KH2PO4 and 5% HEC) at 37 degree Celsius for 5 days. The appliance was kept in a humidified environment when outside the mouth.3%) (p<0. and 27.07 ± 14.6% ± 1. Yuan Y. This was repeated every day for seven days. Method: The study utilized a double blind. using an in situ remineralization model. IADR.01).01). Remineralisation by chewing gum containing CPP-ACP and citric acid Cai F. placebo creme without 2 CPP-ACP and fluoride. Ten subjects were instructed to wear removable palatal appliances. 1. with 4 half-slab insets of human enamel containing demineralized subsurface lesions.GC MI Paste – Abstract booklet – October 2007 89. acid challenged and control) were then embedded. crossover design with three treatments: (i) sugar-free gum (2 pellets) containing 18. IADR. GC Corporation. Conclusion: This study demonstrated significant subsurface enamel remineralization by CPP-ACP tooth creme in the mouth. creme containing 10% CPP-ACP plus 900ppm Fluoride (TMP).Abstract 190 Objectives: Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) have been shown to remineralize enamel subsurface lesions in situ. Remineralization potential of CPP-ACP and its synergy with fluoride Sakaguchi Y.amorphous calcium phosphate (CPP-ACP) in a tooth creme and its synergy with fluoride. TM).5 x 1 mm ) on polished bovine 3 tooth slabs (10 x 5 x 1 mm ) were created by exposure to a lactic acid gel (0.Abstract 191 Objectives: To evaluate the remineralization potential of casein phosphopeptide . Sato T. Nagao S.2%) than chewing with either the gum containing no CPP-ACP or citric acid (9. 14d). The 16hr acid challenge of the remineralized lesions showed that the level of mineral after acid challenge was significantly greater for the gum containing CPP-ACP and citric acid when compared with the other two gums (p<0. and (iii) sugar-free gum not containing CPP-ACP or citric acid. The slabs were then mounted in a buccal flange on a removable maxillary appliance (4 lesions/appliance).1% ± 2. At the completion of each treatment the enamel half-slabs were removed and half of the remineralized lesion treated with carbopol/lactic acid for 16hr. th 49 . Reynolds EC. Results: Chewing with the gum containing CPP-ACP and citric acid resulted in significantly higher remineralization (13. and a professional paste containing 950 ppm fluoride.17.80. The lesions were subjected to analysis using an X-Ray CT scanner (TOSCANER-30000mhd. Enamel subsurface lesions (2. fluoride paste. Subjects were instructed to brush their teeth after lunch with a fluoride toothpaste (900 ppm).13. Conclusion: Sugar-free chewing gum containing CPP-ACP and citric acid significantly promoted remineralization of enamel subsurface lesions in situ. and the acid resistance of the remineralized enamel. 84 General Session. Shen P. randomized. The enamel slabs (remineralized. and to chew gum (2 pellets) for 20 min (4times/d. Kariya S.1 M. and keep the appliance in the mouth for four more hours.8 mg CPPACP and 20 mg citric acid.01).76 ± 10. TMP was 5.3% ± 1.1 with 3 mM CaCl2. 12. sectioned and subjected to microradiography to determine the level of remineralization. Brisbane 2006 . with a synergistic effect of CPP-ACP and fluoride. cover the lesions with the tooth creme. 84 General Session.

IADR. Devani C.3±17.4±16. CDH scores were reduced significantly at 4wk (on average by 46. Results: The 0. o 1.5 and 1% CPP-ACFP solutions both produced statistically similar levels of remineralization as measured by TMR and QLF respectively. 84 General Session.8 and 64.0%w/v) at pH 5. At each of the 4wk and 6 wk time points.GC MI Paste – Abstract booklet – October 2007 91. Objectives: The aim of this study was to examine the remineralization potential of two concentrations of CPP-ACFP in terms of mineral and visual change. fluoride.0 1% 58. Methods: Enamel subsurface lesions were created using the Carbopol method and remineralized using CPP-ACFP solutions (0. there were no significant differences between the two treatment groups. respectively) and at 6wk (by 56. The lesions were then digitally photographed. 0. or that reduce the excitability of pulpal nerves.01 at 4wk and P<0. Brisbane 2006 . both CPP-ACP gel and the KNO3 dentifrice give similar useful reductions in self-rated symptoms of CDH. oxygen and chlorine were collected across the lesions using wavelength dispersive spectrometry on a JEOL 8900 SuperProbe Microprobe. Elemental maps and quantitative line scans for calcium.Abstract 947 The clinical problem of cervical dentinal hypersensitivity (CDH) can be managed by strategies that occlude patent dentine tubules exposed to the oral environment.6 P value 0. Cakar A. Objectives: This randomized clinical trial compared the therapeutic effect of a 10% CPPACP gel (GC Tooth Mousse) with a well established KNO3 dentifrice (Colgate Sensitive) over 6 weeks. Reynolds EC. Conclusion: CPP-ACFP solutions remineralized enamel subsurface lesions in vitro by the deposition of fluorapatite increasing mineral content and improving translucency. embedded.5. and each of these had relatively low baseline scores. IADR. Results: The two groups were well matched with no significance difference in baseline scores. Hall A.85 0.1±6. QLF and TMR analysis of CPP-ACFP remineralized enamel in vitro Cochrane NJ. only 2 of the 18 subjects did not show a response.9% and 46.36 th %R %Q All white spot lesions became more translucent after remineralization treatment.5 (2mL. Effect of CPP-ACP versus potassium nitrate on cervical dentinal hypersensitivity Walsh LJ. th 50 .Abstract 192 Casein phosphopeptides (CPP) have been shown to stabilise amorphous calcium phosphate (CPP-ACP). when assessed using a repeated measures ANOVA (P<0. n=10 lesions per group). Conclusions: Despite differences in their apparent mechanisms of action.8%. Data were analyzed in a blinded manner. The mineral formed was subsequently analysed using wavelength dispersive microprobe spectrometry. In both the CPP-ACP and KNO3 groups. 10 days.7±5. 92. The further reduction in CDH scores which occurred from weeks 4-6 in both groups was not statistically significant.4%.5% lesions. 84 General Session.9 93. Brisbane 2006 . or the KNO3 dentifrice twice daily. Microprobe analysis of remineralized enamel showed a single calcium phosphate phase with the ratio matching apatite. Preliminary studies have shown that topical application of a CPPACP gel can cause blockage of dentine tubules. Cai F. Tran KK. daily solution changes.7 83. sectioned. Wang WS. CPP has also been shown to stabilise amorphous calcium fluoride phosphate (CPP-ACFP) which had an additive effect compared with the separate remineralization effects of fluoride or CPP-ACP.001 at 6wk). Fluoride was found evenly throughout the body of the lesions supporting fluorapatite formation. phosphorous. and a composite score calculated. lapped and analysed using transverse microradiography (TMR) to determine percentage mineral change (%R). however there was more variation in the 0.5% 56. respectively). Methods: Patients (n=18 per group) screened from a general practice setting presenting with CDH used either CPP-ACP gel applied topically each night before retiring in conjunction with their conventional dentifrice twice daily. In each group. 37 C. CDH responses to 4 types of stimuli were selfrated using a visual-analogue scale after 4wk and 6wk of treatment. analysed using quantitative light induced fluorescence (QLF) to determine percentage fluorescent loss integrated over area (%Q). CPP-ACP has been associated with anticariogenicity and remineralization in in vitro and in vivo models. when compared with their relevant baseline values.

the oral environment. Effect of CPP-ACP on hardness of enamel eroded by Cola-drink Sukasaem H. 210. Ca and Pi release.2).7±3.Abstract 1269 Plaque mineralisation is a highly complex multi-factorial process influenced by microbial promotors and inhibitors. group 3 (248. When plaque biofilms were mineralised with CPP-ACP and CPMU together. Pi and F. Brisbane 2006 . Different mechanisms of mineralisation may be involved with more complex pH-dependent CPP-ACP accumulation. Results: The hardness of the specimens at baseline. depending on application regime but under otherwise similar environmental conditions of growth and pH.6±12.2.6. th 51 . continuously supplied (3.2. Poolthong S. The CPP-ACP and CPP-ACP with artificial saliva groups demonstrated significantly higher hardness value than the artificial saliva group (p<0. Results: Mineralisation with CPP-ACP yielded Ca and Pi levels 20-50% that of CPMU. After remineralization.9±7.Abstract 1673 Objective: This in vitro study evaluated the remineralization effect of Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP) from Toothmousse (GC Asia Dental. Plaque pH is a key regulator of plaque mineralisation with CPP-ACP as well as CPMU. Methods: Ten extracted human premolars were bucco-lingually cut into 2 halves and embedded in resin providing 20 specimens with buccal and lingual experimental sites.2. 84 General Session.Ltd. and examine the effect of pH and the presence of F.6±6.1±13..GC MI Paste – Abstract booklet – October 2007 93. Sucrose (5%) was supplied periodically (1. Conclusions: CPP-ACP was effective in mineralising plaque microcosms but mineral deposition was less than with CPMU.8.05). were measured. After remineralization process. 242. and (ii) CPMU (20mM Ca. Brisbane 2006 . Thailand) (5s each for 10 cycles) for 3 times with 2 immersions in artificial saliva for 6h among the 3 alternate immersions. Objective: To deposit caries-protective minerals in microcosm plaque biofilms using (i) casein phosphopeptide-amorphous calcium phosphate complex (CPP-ACP) which models salivary CaPi carriers. CPP-ACP and artificial saliva (Group 3) and deionized water as control. 207. A raised plaque pH range yielded greater CaPi deposition from CPP-ACP.0) mineralising solution which deposits CaPi following a urea-induced pH rise. all groups showed significantly higher hardness value than control (p<0. 12mM phosphate.1±9. group 2 (245.3±5.05 ppm) in DMM. Supported by Health Research Council of New Zealand and Wellington Medical Research Foundation.1±17. A sucrose-induced low pH during CPP-ACP exposure appeared to lower CaPi accumulation. artificial saliva (group 2).5.3±6. Statistical analysis used Oneway ANOVA followed by Bonferroni test (alpha = 0. IADR. Sissons CH. The demineralization process was done by immersion of specimens in Cola-drink (Thai Namthip Co. 8hrly). Deposition from CPP-ACP also increased considerably in the presence of a low concentration of fluoride (0. after demineralization and after remineralization of group 1 (246. For remineralization process.5ml/6min. and from CPMU. 205.2±6. The remineralization effect of CPP-ACP is significantly higher than that of artificial saliva in vitro. Microcosms were mineralised under continuous or periodic regimes. there was a large increase in CaPi deposited. Methods: Plaque microcosms were grown for 11-21days from plaque-enriched saliva in a multi-plaque ‘artificial mouth' culture system (MAM). Plaque pH. 228. and subsequent deposition as CaPi mineral.6ml/hr/plaque) with nutritional saliva analogue (BMM) or chemically-defined artificial saliva (DMM).9. the demineralized specimens were randomly divided into 4 groups (n=10) and 4 regimens of remineralization used CPP-ACP (group 1). and plaque pH.4. IADR. Plaque microcosm biofilm mineralization by CPP-ACP and calcium-phosphate-monofluorophosphate-urea mineralising solution Wong L.05).05).5) were demonstrated with standard deviation respectively in bracket. hardness measurements were repeated. Thailand) and artificial saliva (Chulalongkorn University. 84 General Session. Conclusion: CPP-ACP from Toothmousse can increase hardness of enamel eroded by Cola-drink. 247. 209. South Africa) and baseline Vickers hardness was measured (100g force. 500mM urea. 5mM monofluorophosphate. Imptech.0±8. th 94. The specimens were polished (1 micron diamond slurry.Panich M. 207. Ca. 15s) to demonstrate the effect of mineral changes. supplemented with urea (or glucose) to raise (lower) the resting pH. pH5.0±8.2). Japan) on hardness of enamel eroded by Cola-drink. All specimens were rinsed with deionized water and the hardness measurements were repeated.1) and control group (239.

95kg with deionised water and HCl (pH=3) as lubricants. and also displayed smoother facets.0 kg with hydrochloric acid lubricant (pH = 1.000 cycles) (p<0. and has also highlighted the importance of lubricants in reducing wear. Ranjitkar S. Further research is required to clarify its clinical usefulness of Tooth Mousse® in this context.000 cycles using a Dr PICZA 3D Scanner (PIX-4) and MATLAB software package (version 6. Brisbane 2006 . USA). but no Tooth Mousse® was applied. and enamel halves of the same teeth were worn against each other in a purpose-built electromechanical tooth wear machine under a load of 10.2) for around 10. and with regular Tooth Mousse® application. Kaidonis JA. Enamel wear prevention under conditions simulating bruxism and acid regurgitation Ranjitkar S. the machine was stopped every two minutes (160 cycles of wear) and the specimens washed and dried. Results: The rate of enamel wear was significantly lower in the experimental sample (0. The control group worn with HCl (pH=3) exhibited less wear than that worn with deionised water. and assessed qualitatively by examining epoxy resin replicas under a scanning electron microscope. th 96. Results: Dentine specimens worn with Tooth Mousse® as the sole lubricant exhibited minimal wear.GC MI Paste – Abstract booklet – October 2007 95. USA). Townsens GC.Abstract 2428 Tooth wear is a growing public health problem and there is a need to better understand its aetiology and management. Enamel wear facets in the experimental sample were also found to be smoother than those in the control sample. and several approaches are being used to manage it. Methods: Eight dentine specimens from the lingual halves of third molar teeth were worn against enamel antagonists under a load of 9. These data were then compared using ANOVA with data from two control experiments conducted under a load of 9. under conditions simulating bruxism and acid regurgitation. 84 General Session.95kg in an electro-mechanical tooth wear machine with hydrochloric acid lubricant (HCl) (pH=3). In the experimental sample (n = 8). Kaidonis JA.000 test cycles.01 mm3 per 1. Tooth wear was quantified by measuring reduction in enamel volume per 1.Abstract 2424 Tooth wear is a significant problem facing clinicians. The specimens were further washed and dried before the cycle was continued. Those worn with HCl (pH=3) with regular Tooth Mousse® application showed less wear than both the control groups. The Mathwork Inc. Brisbane 2006 . Natick MA.41 mm3 per 1. Conclusions: This study has shown that Tooth Mousse® is capable of reducing dentine wear.000 cycles) than in the control sample (1. The same protocol was followed for the control specimens (n = 8) but no Tooth Mousse was applied. Objective: This study aims to quantitatively and qualitatively test the efficacy of Tooth Mousse® in managing dentine wear under highly controlled conditions. An in vitro study of wear prevention in dentine Narayana T. IADR.01). Townsend GC. 84 General Session. A further eight dentine specimens were worn with Tooth Mousse® as the sole lubricant. Their wear facets were not obviously different from facets of control specimens worn with HCl (pH=3). Objective: Our aim was to investigate the effectiveness of frequent applications of Tooth Mousse (GC Corporation) in preventing enamel wear in vitro. th 52 . The Mathwork Inc. but were smoother than those of specimens worn with deionised water as the lubricant. Richards LC. Tooth Mousse was then applied for four minutes. These findings open up new possibilities for the prevention of tooth wear. Wear rates were compared between the samples with an unpaired t-test. Qualitative assessment was also carried out using Scanning Electron Microscopy (SEM). Natick MA. IADR. and had very smooth and shiny wear facets. Richards LC. Dentine wear was quantified by measuring reduction in dentine volume using a Dr PICZA 3D Scanner (PIX-4) and a MATLAB software package (version 6. Methods: Sixteen human third molar teeth were sectioned longitudinally in a mesio-distal direction. probably due to its lubrication properties. Conclusions: Frequent application of Tooth Mouse is effective in reducing enamel wear under conditions simulating bruxism and acid regurgitation.

with 1 session supervised on school days.0 in the case of casein phosphopeptide β-casein(1-25). th 53 . Reynolds EC.Abstract 2445 Objectives: To investigate the radiographic progression and regression of dental caries in adolescent subjects chewing a gum containing CPP-ACP over a two-year period. The CPP-ACP gum enhanced regression of carious lesions compared with the control gum.923 ± 0.526 ± 0. Tsao CE. Structure and 15N-dynamics of casein phosphopeptide-amorphous calcium phosphate nanocomplexes Cross KJ.4mg CPP-ACP significantly slowed progression and enhanced regression of dental caries in a two-year clinical trial relative to a normal sugar-free gum. were assessed for approximal surface dental caries at both the enamel and dentine level.044 nm at pH 6. A greater percentage of approximal surfaces remained unchanged with the CPP-ACP gum than with the control gum. The CPP-ACP gum slowed progression of carious lesions compared with the control gum. Brisbane 2006 . Standardised digital radiographs were taken at the baseline and at the completion of the clinical trial using the Dexis digital X-ray system. Features of the model suggest a mechanism by which calcium ion transport through biological membranes and the formation of large CPP-ACP aggregates can be achieved. Results: A molecular model of the CPP-ACP nanocomplex was developed. th 98. All subjects received accepted preventive procedures.41%) of approximal surfaces experienced caries progression compared to 932 (5. Huq NL. scored by a single examiner. Methods: 2720 subjects were randomly assigned to either a test or control group.31%) approximal surfaces in the control group. over the 2-year study period. 814 (4.001). These complexes act as biological calcium phosphate delivery vehicles and are effective in the remineralization of early enamel lesions. Analysis of caries progression or regression was undertaken using a transition matrix. Science and Technology. Conclusion: The CPP-ACP nanocomplexes were shown to be particles with hydrodynamic radii ranging from 1. Attard TJ. IADR. Methods: NMR spectroscopic techniques were used to characterize translational diffusion of the CPP-ACP nanocomplexes and to characterize interatomic distances and the nuclear relaxation behaviour of the peptides within the CPP-ACP nanocomplex.0 increasing to 1. fluoridated dentifrice. For subjects chewing the CPP-ACP gum. Conclusion: A chewing gum containing 54. Supported by NH&MRC grant 209042 .GC MI Paste – Abstract booklet – October 2007 97. Adams GG. Results: There was a statistically significant difference in the distributions of the transition scores between the two groups (P value < 0.Abstract 2534 Tryptic phosphopeptides derived from milk caseins are known to associate with amorphous calcium phosphate (ACP) forming stable complexes. IADR. Study Sponsor: Cadbury Schweppes. Subjects were instructed to chew their assigned gum for 10 minutes 3xday.21%) approximal surfaces with the control gum. including fluoridated water. a reduction of 16. Aim: To examine the structures of these complexes. 56 (0. 84 General Session.30%) of approximal surfaces experienced caries regression with the CPP-ACP gum compared to 36 (0. Reynolds EC. CPP-ACP gum slows progression and enhances regression of dental caries Morgan MV. Brisbane 2006 . 84 General Session. and access to professional care. Bailey DL. The test group received a sugar-free gum containing 54.4mg CPP-ACP while the control group received an identical gum without CPP-ACP. The radiographs.9%.082 nm at pH 9.

Walker GD. Improved plaque uptake and enamel remineralization by fluoride with CPP-ACP Reynolds EC. (iv) 2% CPP-ACP and (v) 2% CPP-ACP plus 1100 ppm F.4%) and the paste containing 2% CPP-ACP plus 1100 ppm F was superior (21. (ii) 1100 ppm F. 54 .Abstract 2538 Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) has been shown to slow the progression of caries and to remineralize enamel subsurface lesions.51 :(4):284-289 The development of white spot demineralization associated with fixed appliance orthodontic treatment is a significant clinical problem. 84 General Session.0±17. Results: The addition of 2% CPP-ACP to the 450 ppm F rinse significantly increased the incorporation of fluoride ions into plaque where the plaque fluoride level (33. IADR. and Toothpastes: (i) placebo. double-blind cross-over studies were designed involving three mouthrinses and five toothpastes as follows: Mouthrinses (i) 2% CPP-ACP. Cai F. Conclusion: CPP-ACP promotes the incorporation of fluoride into plaque and sub-surface enamel producing effects superior to fluoride alone.5%±1. (ii) 2% CPP-ACP plus 450 ppm F and (iii) 450 ppm F. Reynolds C. The toothpaste containing 2% CPP-ACP produced a level of remineralization (13. Objectives: The aim of the studies was to determine the ability of CPP-ACP to increase the incorporation of fluoride into supragingival plaque and to promote enamel remineralization in situ with acid resistant mineral. (iii) 2800 ppm F.6 nmol/mg dry wt) was over double that obtained with the fluoride-only rinse (14. The mouthrinses (15 ml) were used for 60 s. Cochrane NJ.5%±2. Prevention of white spot lesions in orthodontic practice: a contemporary review Sudjalim TR. Both established and experimental methods for prevention of such lesions in day-to-day clinical practice are presented and discussed. Brisbane 2006 . Manton DJ. Australian Dental Journal 2006.0%±5.9%) to all other formulations in enamel lesion remineralization. The toothpastes (1 g) were added to 4 ml water to form a slurry and used for 60 s four times per day for 14 days in an in situ remineralization model. th 100.GC MI Paste – Abstract booklet – October 2007 99.4±6. Woods MG. Methods: Randomized. Shen P. Acid challenge of the remineralized lesions showed that the CPP-ACP/F mineralized lesions were relatively acid resistant.5%) similar to the 2800 ppm F paste (15. three times per day for 5 d and supragingival plaque collected and analyzed for F content. Morgan MV. Fluoride in the toothpaste slurry produced a dose-response related remineralization of subsurface enamel lesions.7 nmol/mg dry wt).

embedded in resin and microradiographic images taken. Incorporation of casein phosphopeptide-amorphous calcium phosphate into a temporary cement Wong R. 3. 84 General Session.9%. 4.0% CPP-ACP delayed it beyond the ISO requirements of 10 minutes.0%. Adams G.4 ± 0.GC MI Paste – Abstract booklet – October 2007 101. TM Increasing amounts of CPP-ACP tended to reduce the compressive strength of Freegenol from a mean of 5 MPa for the 0. double-blind cross-over study was conducted. In situ remineralisation by sugar-free gums. Tokyo. Cai F.0% w/v CPP-ACP into Freegenol would require the addition of an accelerator. Ten healthy adult subjects were recruited. Control specimens contained no CPP-ACP. compressive strength and film thickness. This study is supported by the NHMRC grant no: 359318 and the Cooperative Research Centre for Oral Health Science. These values were in compliance with ISO recommendations of 35 MPa as the maximum for compressive strength. Trident White therefore produced TM TM 107% more remineralisation than the Orbit gum and 75% more than the Orbit Professional . A randomized. Results: Addition of ≤CPP-ACP reduced the setting time but ≥3. Japan) were weighed to a ratio of 1:4 and mixed with 0.0%. with Or 8.5% and OrP 10. September 2005 – Abstract 0020 Objectives: To compare the efficacy of a CPP-ACP containing sugar-free chewing gum TW (Trident White) with sugar-free gums not TM TM containing CPP-ACP: Or (Orbit ) and OrP (Orbit Professional) to remineralise enamel subsurface lesions (WSL) in an in situ model. Walker GD.0% CPP-ACP into Freegenol is otherwise viable in terms of its compressive strength and film thickness. Brisbane 2006 . Methods: Specimens for WSL mineralization were sectioned from either buccal or lingual surfaces of extracted sound third molar teeth.5 ± 0. Percentage remineralisation (%R) was calculated according to the formula (1.ΔZt/ ΔZc ) x 100. The addition of up to 8. Conclusion: The addition of CPP-ACP to sugar-free chewing gum significantly increased the mineralization of WSL in situ when compared to two other gums not containing CPP-ACP. Messer LB. Enamel half slabs were paired with their control. 45 Annual Meeting of Australian/New Zealand Division of the IADR. 2. Test conditions were conducted using ISO recommendations for setting time.0% test group. Cochrane N. with the difference in the test specimen designated ΔZt. Reynolds EC. and the WSL were created using the carbopol method. Subjects chewed the gum for a 20 min period 4 times per day for 14 days. due to its effect of TM delaying setting time beyond ISO recommendations.9%. Reynolds C. This study was carried out to determine the viability of incorporating CPP-ACP into a zinc oxide non-eugenol temporary cement in terms of its effect on setting time.5% CPP-ACP to 3 MPa for the 8. 1. TM Methods: The base and catalyst pastes of Freegenol (GC Int Corp. one-containing CPP-ACP Manton DJ. Shen P.0%. School of Dental Science.5%. The difference in mineralization profile in the control specimen between the sound and WSL enamel windows was designated ΔZc. Wilson PR.0% and 8. TM th 102. TM Results: TW produced remineralisation (%R) of 18. This study was supported by the Cooperative Research Centre for Oral Health Science. TM Conclusion: The incorporation of ≥3.Abstract 0653 Objectives: The presence of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) in a temporary cement may act as a surface modifying agent which alters the permeability of cut dentin deneath temporary crowns. IADR.0% w/v of CPP-ACP powder. Five specimens were used in each test group. th 55 . Palamara J. The University of Melbourne.9 ± 0. All differences were statistically significant. Film thickness measurements for all test groups complied with the recommended maximum of 25µm. compressive strength and film thickness for Type 1-Class 1 zinc exide non-eugenol cements.

56 . Japan) on its physical properties. a significant increase in sonic velocity was found for specimens stored in the CPP-ACP solution.1M lactic acid buffer solution (pH4. Miyazaki M.05) were done. Panametrics). IADR. Tokyo. Results were analyzed using one-way ANOVA and the Kruskal-Wallis tests. It could be concluded that the inorganic components contained in high concentrations in CPP-ACP acted to enhance remineralization of the enamel structure. Results: The effect of incorporating up to 10% CPP-ACP into Fuji III on the values of ST. This project is supported by The Cooperative Research Centre for Oral Health Science. Palamara J. Takamizawa T. 3. DTS and FV remained within ISO recommendations. Results: The sonic velocity was found to decrease with time for specimens stored in the demineralization solution.0). Effect of CPP-ACP paste on mechanical properties of bovine enamel as determined by an ultrasonic device Yamaguchi K. in addition to a prolonged ST. The values of CS and DTS decreased with a significant effect when 10% CPP-ACP was added. P 0. 7 and 10% CPP-ACP were tested following the international Organization for Standardization (ISO) specifications 9917:1991 and 6876:1986. 1. Conclusion: From the result of this study.and remineralization of the enamel structure could be measured non-destructively by using an ultrasonic pulse method. Burrow M. Methods: The enamel specimens were prepared by cutting bovine teeth into blocks. Diametral Tensile Strength (DTS). Film Thickness (FT) and Flow values (FV) of Fuji III containing 0. and then stored in the artificial saliva (pH 7. and one-way ANOVAs followed by the Tukey HSD tests (α=0. Messer H. Moore BK. On the other hand. Ca 0. Incorporation of casein phosphopeptide-amorphous calcium phosphate into glass ionomer cement Al-Zraikat H. Conclusion: There was no adverse effect following the incorporation of up to 10% CPP-ACP on the physical properties tested except on FT with 5% CPP-ACP. Other specimens were stored in a 10-times diluted solution of CPP-ACP paste and a placebo paste without CPP-ACP for 10 min. 5. Six specimens were used for each condition. th 104. The specimens were stored in 0. Fuji III containing 5% CPP-ACP failed the FT test..Abstract 0654 Objectives: The incorporation of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) into glass ionomer cement (GIC) fissure sealant may enhance the inhibition of demineralization and promote remineralization. it was suggested that the conditions of de. The aim of this study is to investigate the effect of incorporating increasing concentrations of CPP-ACP into Fuji III fissure sealant (GC Int. Compressive Strength (CS). 84 General Session.75mM.GC MI Paste – Abstract booklet – October 2007 103.45mM) for 10 min twice a day. Methods: Setting time (ST). CS.75. followed by 10 min immersion into a demineralization solution twice a day before storage in the artificial saliva. Journal of Dentistry (2006) 34. 230-236 Objective: The purpose of this study was to determine the effect of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste on demineralization of bovine enamel by measuring changes in the ultrasound transmission velocity. Corp. Brisbane 2006 . Inage H. The propagation time of longitudinal ultrasonic waves was measured by a Pulser-Receiver (Model 5900. Panametrics) with a transducer (W112.

to enamel treated with cerbamide peroxide and/or CPP-ACP were investigated. The sonic velocity of the demineralized specimens was found to decrease significantly over time. each being allocated into one of the four groups (n=26): bleach (Polanight.05). Hurokawa H. Conclusion: These findings suggest the shear bond strength of resin to enamel using a self-etching priming adhesive may be affected if the enamel is treated with a bleaching agent or CPP-ACP. 41:204-207 The purpose of this study was to investigate the demineralization of dentin by measuring changes in the velocity of the sonic longitudinal waves transmitted through this substrate. Only combined use of carbamide peroxide and CPP-ACP significantly affected microshear bond strength with Single Bond. Angelis F. 16% carbamide peroxide). SE Bond showed the highest bond strength to untreated enamel (p<0. 522):133-137 Background: Limited evidence exists regarding the effect of carbamide peroxide and casein phosphopeptide amorphous calcium phosphate (CPP-ACP) on composite-enamel bonding. 57 . Resin bonding using an all-etch or self-etch adhesive to enamel after carbamide peroxide and/or CPP-ACP treatment Moule CA. Inage H. The microshear bond strength of SE Bond decreased when the enamel was treated with carbamide peroxide. Microshear bond strengths. No significant increase in sonic velocity was observed in specimens treated with CPP-ACP. Foo MS.Australian Dental Journal 2007. One group of samples was immersed in demineralization solution for 10 min twice a day and then stored in artificial saliva. Thomas D. CPP-ACP or both (p<0. or untreated (control). Materials & methods: Twenty-six extracted human third molars were sectioned into four parts. CPP-ACP (GC Tooth Mousse).GC MI Paste – Abstract booklet – October 2007 105. Miyazaki M. Results: A significant difference in bond strength was found between bonding systems. Takamizawa T. Burrow MF. Malipatil S.05). 106. bleach and then CPP-ACP. Kim GH. using either a total-etch or self-etching adhesive. Le S. Two additional groups of samples were treated with a solution of casein phophopeptide-amorphous calcium phosphate (CPP-ACP) paste or a placebo paste without CPP-ACP before demineralization and a control group was stored in artificial saliva. Caries Res 2007. The surfaces were bonded with a total-etch bonding system (Single Bond) or a self-etching primer system (Clearfil SE Bond) and tested using a microshear test. suggesting that CPP-ACP acted to prevent demineralization. Ultrasonic determination of the effect of casein phosphopeptide-amorphous calcium phosphate paste on the demineralization of bovine dentin Yamaguchi K.

Yamaguchi K. and were observed under field emission-scanning electron microscopy (FE-SEM). Vougiouklakis G. On the other hand. Inage H. 115-120. they were fixed in 2.5% glutaraldehyde in cacodylate buffer solution. The SEM observations revealed different morphological features brought about by the various storage conditions. have been shown to exhibit anticariogenic properties in human and animal models.1 M lactic acid buffer solution for 10 min and then in artificial saliva (negative control). The specimen surfaces were subjected to surface analysis by Fourier transformance micro multiple internal reflectance infrared spectroscopy (micro MIR-FTIR). Irokawa A. Vol. Takamizawa T. C and D (n=10). The remaining specimens were stored in a 10 times-diluted solution of CPP-ACP paste or a placebo paste containing no CPP-ACP for 10 min. The specimens were prepared by cutting enamel and dentin of bovine teeth into blocks. B. Effect of CPP-ACP paste on tooth mineralization: an FE-SEM study Oshiro M. Journal of Oral Science. Group C resulted in a significant higher %RM compared to group D.49. 2007 Milk and milk products. Watanabe T. The specimens were divided in four groups the A. From the morphological observations of the enamel and dentin surfaces.2. Effect of a CPP-ACP agent on the demineralization and remineralization of dentine in vitro Rahiotis C.75. Miyazaki M. such as cheese. dehydrated in ascending grades of test-butyl acohol. Watanabe T. while no agent was applied on specimens of group D. Afterwards. Conclusions: The presence of agent CPP-ACP on dentine surfaces provoked lower demineralization and higher remineralization in comparison with the dentine surfaces without agent. CPP-ACP shows an anti-caries effect by suppressing demineralization. C and D were immersed in demineralization solution for 7 days. 21 and 28 days. while no agent were applied on the specimens of B group. Ca) twice a day before storage in artificial saliva. groups A. After treatment of the specimens for 3. or possibly a combination of both.GC MI Paste – Abstract booklet – October 2007 107. enamel and dentin specimens treated with CPP-ACP paste revealed slight changes in their morphological features. enhancing remineralization. Takamizawa T. Tooth mousse was applied on surface specimens of A group. and then transferred to a critical-point dryer. The surfaces were coated with a thin film of Au in a vacuum evaporator. the surfaces were subjected to micro MIR-FTIR analysis and the mineral to matrix ratio was used to assess the extent of dentin demineralization (DM). Yamaguchi K. 7. Results: Group A showed significant lower %DM in comparison to group B. Journal of Dentistry 35 (2007) 695-698 Objectives: The aim of this study was to determine in vitro the effect of a commercial paste based on CPP-ACP complex on the demineralization of sound human dentine and on remineralization potential of artificial caries-like lesions formed on dentine surfaces. it could be considered that the CPP-ACP paste might prevent demineralization of the tooth structure. B. Inage H. The purpose of this study was to evaluate the effect of CPP-ACP paste on demineralization by observing the treated tooth surface using an FE-SEM. Methods: Forty dentine specimens were prepared with hard tissue microtome. Tooth mousse was applied on specimens of group C. followed by 10 min immersion in a demineralizing solution (pH=4. The groups C and D immersed in artificial saliva for 7 days and were subjected to analysis by micro MIR-FTIR and the mineral to matrix ratio was used to assess the extent of dentin remineralization (RM). Ando S. No. A few specimens were stored in 0. Afterwards. Irokawa A. Miyazaki M. 108. Demineralization of the enamel and dentin surfaces was more pronounced with the longer test period in the control and negative control specimens. 58 . Ando S.

placebo-controlled. The enamel slabs were removed from the appliance. GC Corporation.0 mM Ca and 1. make teeth stronger. In the current in situ study. Although more research is needed to assess the broader implications of the technique.1 and 20. Toshiba IT & Control System Corp. additional efforts may be required. Compendium. May 2007. Remineralization of enamel lesion by a novel cream with both CPP-ACP and fluoride Kariya S.6 respectively. 59 . Especially. rinse the lesions with water. Enamel slabs with unpolished surfaces (2x3x4mm) were exposed to a lactic acid buffer gel (0.6±8. MIPP showed higher remineralization potential than the FP in this test (p<0. cut into thin sections of approximately 200µm thickness. mean age 25y) were recruited.8mM P) at 37 degree Celsius for 5 days to create artificial enamel sub-surface lesions.3±7.05). The patient’s teeth successfully lightened from an initial shade of A2 to a uniform shade of B1. This clinical case demonstrates the use of a combination in-office and take-home whitening regimen. Kato S. Results: The mean percentage remineralization for the PP. 14. crossover design study. Conclusions: This in situ study confirmed the remineralization effect of CPP-ACP and demonstrated a synergistic effect of CPP-ACP and Fluoride. and help protect them from decay and erosion. When it is insert into the mouth.1M. These results indicate that the CPP-ACP and Fluoride (NaF) can coexist and remineralization potential of fluoride is further enhanced by CPP-ACP.5 with 3. Iijima Y. The appliance was stored in a humidified environment whilst not in the mouth. randomized. followed by the in-office and home application of a paste containing casein phosphopeptide and amorphous calcium phosphate. 110. and keep the appliance in the mouth for a further four hours. Discussion. one third of each enamel slab was covered by nail varnish as a negative control. The white spots that were still visible after the bleaching process had disappeared into the tooth structure after the 3-week at-home use of the remineralizing paste. Sakaguchi Y. two enamel slabs are placed adjacent to the buccal surface of mandibular molars. Extracted sound human first premolar teeth were obtained from the dental hospital of Nagasaki University. Poster session 136 – 54th Annual ORCA Congress 2007 Objective: The aim of this two-center. Japan) Methods: This was a two-center. Japan) and Scion Image for Windows (Scion Corporation). Experimental materials are shown in the table 1. The figure 1 is a photo of the device. pH4. at the 3-month recall appointment. This was repeated every day for 7 days. After demineralization. Volunteers were divided randomly into 3 groups (Table 2).GC MI Paste – Abstract booklet – October 2007 109. in some cases involving stark with spots. The microsections were analyzed using X-Ray µ-CT (TOSCANER-30000µhd. the author concludes that this individualized result suggests that products with the potential to remineralize tooth structure might be useful adjuncts to traditional tooth-bleaching processes to achieve a uniform appearance when white spots are present. Fifteen healthy adults (8 males and 7 females. cover the lesions with the test materials. However. some relapse in the appearance of the white spots was noted. MIPP and FP showed remineralization of enamel sub-surface lesions. randomized trial is to evaluate the remineralization potential of a novel cream with both Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP) and Fluoride (GC MI Paste Plus / Tooth Mousse Plus. 28(5):234-240 Although in-office and dentist-monitored vital night-guard bleaching have proved successful for whitening teeth. both of which are believed to help replace lost minerals in teeth. Sato T. Considering biomodification and remineralization techniques as adjuncts to vital tooth-bleaching regimens Milnar FJ. FP and MIPP was 3.2±7. wear the appliance for 30 min. Ethical approval was obtained from the University of Nagasaki Ethics Committee. They were instructed to brush their teeth after lunch with fluoride free toothpaste.5.. Chen L.

Further improvement of aesthetics was achieved with composite veneers. number 8. Quintessence International volume 38. Krejci I. The proposed technique is based on reactivation of enamel by elimination of its hypermineralized external layer through microabrasion. Aesthetic management of severely fluorosed incisors in an adolescent female Ng F.GC MI Paste – Abstract booklet – October 2007 111. Conclusions: Conservative treatment options such as tooth whitening and microabrasion can dramatically improve severely discoloured fluorosed teeth. Australian Dental Journal 2007. Castioni NV. microabrasion and take-home whitening.523):243-248 Background: Dental fluorosis is a condition of enamel hypomineralization due to the effects of excessive fluoride on ameloblasts during enamel formation. producing hypomineralized and porous enamel. Results: Dark brown to black staining of the teeth was reduced successfully without the need for gross mechanical preparation of the enamel. 112. This can provide a satisfactory interim outcome or minimize the removal of discoloured enamel and dentine prior to the provision of composite veneers. Minimally invasive treatment of white spot enamel lesions Ardu S. September 2007 This article describes a technique used to treat superficial white spot lesions by a minimally invasive approach. The use of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) may enhance remineralization and decrease post-operative sensitivity following tooth whitening and microabrasion procedures in hypomineralized teeth. Methods: A 13 year old girl presented with severely discoloured maxillary central incisors. 60 . Initial aesthetic management of these teeth was conservative. Delayed degradation of enamel matrix proteins or inhibited protein removal results in impaired and incomplete crystal growth. Benbachir N. Manton DJ. followed by daily home application of casein phosphopeptide-amorphous calcium phosphate complexes (CPP-ACP). including in-office tooth whitening. Microabrasion followed by long-term daily home application of CPP-ACP may be considered an interesting alternative to the restorative approach for treatment of white spot lesions. Severely fluorosed teeth may undergo post-eruptive surface breakdown and posteruptive dark brown to black staining. The technique may allow elimination of white spot lesions without involving restorative procedures.